Alkynyl pyrrolo[2,3-d]pyrimidines and related analogs as HSP90-inhibitors

ABSTRACT

Alkynyl pyrrolo [2,3-d] pyrimidines of Formula I are described and demonstrated to have utility as Heat Shock Protein 90 (HSP90) inhibiting agents used in the treatment and prevention of various HSP90 mediated disorders. Methods of synthesis and use of such compounds are also described and claimed.

CROSS-REFERENCE

This application is a division of U.S. patent application Ser. No.11/277,918, filed Mar. 29, 2006, now U.S. Pat. No. 7,544,672, whichclaims the benefit of U.S. Provisional Application No. 60/666,899 filedMar. 30, 2005, all of which are herein incorporated by reference intheir entirety.

FIELD OF THE INVENTION

The invention relates in general to alkynyl pyrrolopyrimidines and theirbroad-spectrum utility, e.g., in inhibiting heat shock protein 90(HSP90) to thereby treat or prevent HSP90-mediated diseases.

BACKGROUND

HSP90s are ubiquitous chaperone proteins that are involved in folding,activation and assembly of a wide range of proteins, including keyproteins involved in signal transduction, cell cycle control andtranscriptional regulation. Researchers have reported that HSP90chaperone proteins are associated with important signaling proteins,such as steroid hormone receptors and protein kinases, including, e.g.,Raf-1, EGFR, v-Src family kinases, Cdk4, and ErbB-2 (Buchner J. TIBS1999, 24, 136-141; Stepanova, L. et al. Genes Dev. 1996, 10, 1491-502;Dai, K. et al. J. Biol. Chem. 1996, 271, 22030-4). Studies furtherindicate that certain co-chaperones, e.g., HSP70, p60/Hop/Sti1, Hip,Bag1, HSP40/Hdj2/Hsj1, immunophilins, p23, and p50, may assist HSP90 inits function (see, e.g., Caplan, A. Trends in Cell Biol. 1999, 9,262-68).

HSP90 possesses a binding pocket at its N-terminus. This pocket ishighly conserved and has weak homology to the ATP-binding site of DNAgyrase (Stebbins, C. et al., supra, Grenert, J. P. et al. J. Biol. Chem.1997, 272, 23843-50). Further, ATP and ADP have both been shown to bindthis pocket with low affinity and to have weak ATPase activity(Proromou, C. et al. Cell 1997, 90, 65-75; Panaretou, B. et al. EMBO J.1998, 17, 4829-36). In vitro and in vivo studies have demonstrated thatoccupancy of this N-terminal pocket by ansamycins and other HSP90inhibitors alters HSP90 function and inhibits protein folding. At highconcentrations, ansamycins and other HSP90 inhibitors have been shown toprevent binding of protein substrates to HSP90 (Scheibel, T. H. et al.Proc. Natl. Acad. Sci. USA 1999, 96, 1297-302; Schulte, T. W. et al. J.Biol. Chem. 1995, 270, 24585-8; Whitesell, L., et al. Proc. Natl. Acad.Sci. USA 1994, 91, 8324-8328). HSP90 inhibitors, e.g. ansamycins, havealso been demonstrated to inhibit the ATP-dependent release ofchaperone-associated protein substrates (Schneider, C. L. et al. Proc.Natl. Acad. Sci., USA 1996, 93, 14536-41; Sepp-Lorenzino et al. J. Biol.Chem. 1995, 270, 16580-16587). In either event, the substrates aredegraded by an ubiquitin-dependent process in the proteasome (Schneider,C. L., supra; Sepp-Lorenzino, L., et al. J. Biol. Chem. 1995, 270,16580-16587; Whitesell, L. et al. Proc. Natl. Acad. Sci. USA 1994, 91,8324-8328).

HSP90 substrate destabilization occurs in tumor and non-transformedcells alike and has been shown to be especially effective on a subset ofsignaling regulators, e.g., Raf (Schulte, T. W. et al. Biochem. Biophys.Res. Commun. 1997, 239, 655-9; Schulte, T. W., et al. J. Biol. Chem.1995, 270, 24585-8), nuclear steroid receptors (Segnitz, B.; U. GehringJ. Biol. Chem. 1997, 272, 18694-18701; Smith, D. F. et al. Mol. Cell.Biol. 1995, 15, 6804-12), v-Src (Whitesell, L, et al. Proc. Natl. Acad.Sci. USA 1994, 91, 8324-8328) and certain transmembrane tyrosine kinases(Sepp-Lorenzino, L. et al. J. Biol. Chem. 1995, 270, 16580-16587) suchas EGF receptor (EGFR) and Her2/Neu (Hartmann, F., et al. Int. J. Cancer1997, 70, 221-9; Miller, P. et al. Cancer Res. 1994, 54, 2724-2730;Mimnaugh, E. G., et al. J. Biol. Chem. 1996, 271, 22796-80(1; Schnur, R.et al. J. Med. Chem. 1995, 38, 3806-3812), CDK4, and mutant p53.Erlichman et al. Proc. AACR 2001, 42, abstract 4474. The HSP90inhibitor-induced loss of these proteins leads to the selectivedisruption of certain regulatory pathways and results in growth arrestat specific phases of the cell cycle (Muise-Heimericks, R. C. et al. J.Biol. Chem. 1998, 273, 29864-72), and apoptosis, and/or differentiationof cells so treated (Vasilevskaya, A. et al. Cancer Res., 1999, 59,3935-40). HSP90 inhibitors thus hold great promise for the treatmentand/or prevention of many types of cancers and proliferative disorders,and also hold promise as traditional antibiotics.

In addition to anti-cancer and antitumorigenic activity, HSP90inhibitors have also been implicated in a wide variety of otherutilities, including use as anti-inflammation agents, anti-infectiousdisease agents, agents for treating autoimmunity, agents for treatingstroke, ischemia, multiple sclerosis, cardiac disorders, central nervoussystem related disorders and agents useful in promoting nerveregeneration (See, e.g., Rosen et al. WO 02/09696 (PCT/US01/23640);Degranco et al. WO 99/51223 (PCT/US99/07242); Gold, U.S. Pat. No.6,210,974 B1; DeFranco et al., U.S. Pat. No. 6,174,875. Overlappingsomewhat with the above, there are reports in the literature thatfibrogenic disorders including but not limited to scleroderma,polymyositis, systemic lupus, rheumatoid arthritis, liver cirrhosis,keloid formation, interstitial nephritis, and pulmonary fibrosis alsomay be treatable with HSP90 inhibitors. Strehlow, WO 02/02123(PCT/US01/20578). Still further HSP90 modulation, modulators and usesthereof are reported in Application Nos. PCT/US03/04283, PCT/US02/35938,PCT/US02/16287, PCT/US02/06518, PCT/US98/09805, PCT/US00/09512,PCT/US01/09512, PCT/US01/23640, PCT/US01/46303, PCT/US01/46304,PCT/US02/06518, PCT/US02/29715, PCT/US02/35069, PCT/US02/35938,PCT/US02/39993, 60/293,246, 60/371,668, 60/335,391, 60/128,593,60/337,919, 60/340,762, 60/359,484 and 60/331,893.

Recently, purine derivatives, including pyrrolopyrimidines showing HSP90inhibitory activity have been reported, e.g., in PCT/US02/35069;PCT/US02/36075. U.S. patent application Ser. No. 10/945,851 andPCT/US04/31248. However, a need remains for additional novel and potentpyrrolopyrimidine HSP90 inhibitors that meet the demanding biologicaland pharmaceutical criteria required to proceed towards human clinicaltrials.

SUMMARY OF THE INVENTION

The present invention is directed towards alkynylpyrrolo[2,3-d]pyrimidines and related compounds that show broad utility,e.g., by inhibiting HSP90 and treating diseases that areHSP90-dependent. These compounds differ from a parent pyrrolopyrimidinewhich was disclosed in prior patent applications in that they aresubstituted with an alkyne, e.g., acetylene, on the ring carbon at thefifth position (C-5 position) and they exhibit improved HSP90 inhibitoryactivity over the parent compounds.

In one aspect, the invention comprises alkynyl pyrrolo[2,3-d]pyrimidinecompounds of Formula I:

wherein:

R⁰ is selected from the group consisting of hydrogen, halogen, loweralkyl, —CN, —SR⁸, —OR⁸, and —NHR⁸;

R¹ is selected from the group consisting of halogen, —OR¹¹, —SR¹¹ andlower alkyl;

R² is —NHR¹;

R³ is selected from the group consisting of hydrogen, —CN, —C(O)OH,—OR¹¹, —SR¹¹, —C(O)R⁹, —NR⁸R¹⁰, lower alkyl, lower alkenyl, loweralkynyl, lower perhaloalkyl, lower alkylsilyl, aryl, heteroaryl,alicyclyl and heterocyclyl, all optionally substituted, wherein:

-   -   the aryl, heteroaryl, alicyclyl and heterocyclyl groups are        mono-, bi- or tri-cyclic;    -   R⁸ and R¹⁰ taken together with the N atom to which they are        attached optionally form an optionally substituted ring        comprising 3-7 ring atoms, wherein, in addition to said N atom,        0-3 of the ring atoms are heteroatoms selected from the group        consisting of O, S and N;    -   the optional substituents on R³ are selected from the group        consisting of lower alkyl, lower alkenyl, lower alkynyl, —CN,        —C(O)OH, —NO₂, —SR⁸, —OR⁸, —C(O)R⁹, —NR⁸R⁸, lower aryl,        heteroaryl, alicyclyl, lower heterocyclyl, arylalkyl,        heteroarylalkyl, amino, alkylamino, dialkylamino,        arylalkylamino, diarylamino, heteroarylamino, diheteroarylamino,        arylheteroarylamino, oxo, perhaloalkyl, perhaloalkoxy,        perhaloacyl, guanidinyl, pyridinyl, thiophenyl, furanyl,        indolyl, indazolyl, phosphonyl, phosphatidyl, phosphoramidyl,        sulfanyl, sulfinyl, sulfonyl, sulphonamidyl, carbamyl, uryl,        thiouryl and thioamidyl, wherein        -   R⁸ and R⁸ taken together with the N atom to which they are            attached optionally form an optionally substituted ring            comprising 3-7 ring atoms, wherein, in addition to said N            atom, 0-3 of the ring atoms are heteroatoms selected from            the group consisting of O, S and N;

R⁴ is selected from the group consisting of optionally substituted loweralkylene, —C(R¹²)₂—, —C(O)—, —C(S)—, —S(O)— and —SO₂—;

R⁵ is selected from the group consisting of aryl, heteroaryl, alicyclyland heterocyclyl, wherein:

-   -   the aryl group is substituted with 2 to 5 substituents;    -   the heteroaryl group is substituted with 2 to 5 substituents;    -   the alicyclyl group is substituted with 3 to 5 substituents;    -   the heterocyclyl group is substituted with 3 to 5 substituents;

the substituents on R⁵ are selected from the group consisting ofhalogen, lower alkyl, lower alkenyl, lower alkynyl, —CN, —C(O)OH, —NO₂,—SR⁸, —OR⁸, —C(O)R⁹, —NR⁸R¹⁰, lower aryl, lower heteroaryl, loweralicyclyl, lower heterocyclyl, arylalkyl, heteroarylalkyl, thioalkyl,amino, alkylamino, dialkylamino, arylalkylamino, oxo, perhaloalkyl,perhaloalkoxy, perhaloacyl, guanidinyl, pyridinyl, thiophenyl, furanyl,indolyl, indazolyl, phosphonyl, phosphatidyl, phosphoramidyl, sulfanyl,sulfinyl, sulfonyl, sulphonamidyl, carbamyl, uryl, thiouryl andthioamidyl, wherein

R⁸ and R¹⁰ taken together with the N atom to which they are attachedoptionally form an optionally substituted ring comprising 3-7 ringatoms, wherein, in addition to said N atom, 0-3 of the ring atoms areheteroatoms selected from the group consisting of O, S and N;

R⁸ is selected from the group consisting of hydrogen, lower alkyl, loweralkenyl, lower alkynyl, lower heteroalkyl, lower heteroalkenyl, lowerheteroalkynyl, lower aryl, lower heteroaryl and —C(O)R⁹;

R⁹ is selected from the group consisting of H, lower alkyl, loweralkenyl, lower alkynyl, lower aryl, lower heteroaryl, —NR¹⁰R¹⁰ and—OR¹¹, wherein

R¹⁰ and R¹⁰ taken together with the N atom to which they are attachedoptionally form an optionally substituted ring comprising 3-7 ringatoms, wherein, in addition to said N atom, 0-3 of the ring atoms areheteroatoms selected from the group consisting of O, S and N;

R¹⁰ is selected from the group consisting of hydrogen, lower alkyl,lower alkenyl, lower alkynyl, lower heteroalkyl, lower heteroalkenyl,lower heteroalkynyl, lower aryl, lower heteroaryl and —C(O)R¹¹;

R¹¹ is selected from the group consisting of lower alkyl, lower alkenyl,lower alkynyl, lower aryl and lower heteroaryl; and

R¹² is selected from the group consisting of hydrogen and lower alkyl.

Also included in the scope of the present invention are stereoisomicforms, including the individual enantiomers and diastereomers, racemicmixtures, and diasteromeric mixtures, and combinations thereof, whereappropriate, as well as polymorphs, specific racemates andstereoisomers, solvates, esters, tautomers, pharmaceutically acceptablesalts and prodrugs of these compounds.

In another aspect, the invention features pharmaceutical compositionscomprising the compounds of the invention, in particular, the compoundsof Formula I, or a polymorph, solvate, ester, tautomer, diastereoisomer,enantiomer, pharmaceutically acceptable salt or prodrug thereof, and oneor more pharmaceutical excipients, for use in treatment or prevention ofdiseases and conditions that are HSP90-dependent.

In yet another aspect, the invention is related to methods of preventingor treating HSP90-mediated disorders or conditions by administering apharmaceutical composition that comprises a pharmaceutically effectiveamount of a compound of Formula I, or a polymorph, solvate, ester,tautomer, diastereomer, enantiomer, pharmaceutically acceptable salt orprodrug thereof.

In one embodiment, the invention provides a method for treating anindividual having a disorder selected from the group of inflammatorydiseases, infections, autoimmune disorders, stroke, ischemia, cardiacdisorders, neurological disorders, fibrogenic disorders, proliferativedisorders, tumors, leukemias, chronic lymphocytic leukemia, acquiredimmuno-deficiency syndrome, neoplasms, cancers, carcinomas, metabolicdiseases, and malignant diseases.

In another embodiment, the invention provides a method for treating anindividual having a fibrogenic disorder, such as, for example,scleroderma, polymyositis, systemic lupus, rheumatoid arthritis, livercirrhosis, keloid formation, interstitial nephritis or pulmonaryfibrosis.

In another embodiment, the invention provides a combination therapycomprising the administration of a pharmaceutically effective amount ofa compound of Formula I, or a solvate, tautomer, diastereomer,enantiomer, pharmaceutically acceptable salt, polymorph, or prodrugthereof according to any of the preceding aspects or embodiments, and atleast one therapeutic agent selected from the group of cytotoxic agents,anti-angiogenesis agents and anti-neoplastic agents. The anti-neoplasticagent may be selected from the group of alkylating agents,anti-metabolites, epidophyllotoxins, antineoplastic enzymes,topoisomerase inhibitors, procarbazines, mitoxantrones, platinumcoordination complexes, biological response modifiers and growthinhibitors, hormonal/anti-hormonal therapeutic agents, andhaematopoietic growth factors.

In a further aspect, the invention is related to the use of thecompounds of Formula I in the manufacture of a medicament.

In yet a further aspect, the invention is related to the use of thecompounds of Formula I in the manufacture of a medicament for thetherapeutical and/or prophylactic treatment of HSP90-dependent diseasesand conditions.

INCORPORATION BY REFERENCE

All publications and patent applications mentioned in this specificationare herein incorporated by reference to the same extent as if eachindividual publication or patent application was specifically andindividually indicated to be incorporated by reference.

BRIEF DESCRIPTION OF THE DRAWINGS

The novel features of the invention are set forth with particularity inthe appended claims. A better understanding of the features andadvantages of the present invention will be obtained by reference to thefollowing detailed description that sets forth illustrative embodiments,in which the principles of the invention are utilized, and theaccompanying drawings of which:

FIG. 1 represents a plot of tumor volume (mm³) against time (days), foranimals administered compounds of the present invention (and controls)in a mouse N87 Gastric Carcinoma Xenograft model, as described inexample 70.

FIG. 2 represents a plot of tumor volume (mm³) against time (days), foranimals administered compounds of the present invention (and controls)in a mouse NC1295 Adrenocortical Carcinoma Xenograft model, as describedin example 71.

FIG. 3 represents a plot of tumor volume (mm³) against time (days), foranimals administered compounds of the present invention (and controls)in a mouse SK-MEL-28 Melanoma Xenograft model, as described in example72.

FIG. 4 represents a plot of tumor volume (mm³) against time (days), foranimals administered compounds of the present invention (and controls)in a mouse a plot of tumor volume (mm³) against time (days), for animalsadministered compounds of the present invention (and controls) in amouse HT29 Colon Carcinoma Xenograft model, as described in example 73.

FIG. 5 represents Western Blot protein analysis of N87 gastric carcinomacells after treatment with compounds 2 and 23 at varying concentrationsand timepoints, as described in example 74.

DETAILED DESCRIPTION OF THE INVENTION

While preferred embodiments of the present invention have been shown anddescribed herein, it will be obvious to those skilled in the art thatsuch embodiments are provided by way of example only. Numerousvariations, changes, and substitutions will now occur to those skilledin the art without departing from the invention. It should be understoodthat various alternatives to the embodiments of the invention describedherein may be employed in practicing the invention. It is intended thatthe following claims define the scope of the invention and that methodsand structures within the scope of these claims and their equivalents becovered thereby.

Definitions

A “pharmaceutically acceptable derivative or prodrug” means anypharmaceutically acceptable salt, ester, salt of an ester or otherderivative of a compound of this invention, which, upon administrationto a recipient, is capable of providing, either directly or indirectly,a compound of this invention or a pharmaceutically active metabolite orresidue thereof. Particularly favored derivatives or prodrugs are thosethat increase the bioavailability of the compounds of this inventionwhen such compounds are administered to a patient (e.g., by allowingorally administered compound to be more readily absorbed into blood) orwhich enhance delivery of the parent compound to a biologicalcompartment (e.g., the brain or lymphatic system).

A “pharmaceutically acceptable salt” may be prepared for any compound ofthe invention having functionality capable of forming a salt, forexample, an acid or base functionality. Pharmaceutically acceptablesalts may be derived from organic or inorganic acids and bases.Compounds of the invention that contain one or more basic functionalgroups, (e.g., amino, alkylamino), are capable of formingpharmaceutically acceptable salts with pharmaceutically acceptableorganic and inorganic acids. These salts can be prepared in situ duringthe final isolation and purification of the compounds of the invention,or by separately reacting a purified compound of the invention in itsfree base form with a suitable organic or inorganic acid, and isolatingthe salt thus formed. Examples of suitable acid salts include acetate,adipate, alginate, aspartate, benzoate, benzenesulfonate, bisulfate,butyrate, citrate, camphorate, camphorsulfonate, cyclopentanepropionate,digluconate, dodecylsulfate, ethanesulfonate, formate, fumarate,glucoheptanoate, glycerophosphate, glycolate, hemisulfate, heptanoate,hexanoate, hydrochloride, hydrobromide, hydroiodide,2-hydroxyethanesulfonate, lactate, maleate, malonate, methanesulfonate,2-napthalenesulfonate, nicotinate, nitrate, oxalate, palmoate,pectinate, persulfate, 3-phenylpropionate, phosphate, picrate, pivalate,propionate, salicylate, succinate, sulfate, tartrate, thiocyanate,tosylate and undecanoate. Other acids, such as oxalic, while not inthemselves pharmaceutically acceptable, may be employed in thepreparation of salts useful as intermediates in obtaining the compoundsof the invention and their pharmaceutically acceptable acid additionsalts. See, e.g., Berge et al. “Pharmaceutical Salts”, J. Pharm. Sci.1977, 66:1-19. Compounds of the present invention that contain one ormore acidic functional groups are capable of forming pharmaceuticallyacceptable salts with pharmaceutically acceptable bases. The term“pharmaceutically acceptable salts” in these instances refers to therelatively non-toxic, inorganic and organic base addition salts ofcompounds of the present invention. These salts can likewise be preparedin situ during the final isolation and purification of the compounds, orby separately reacting the purified compound in its free acid form witha suitable base, such as the hydroxide, carbonate or bicarbonate of apharmaceutically acceptable metal cation, with ammonia, or with apharmaceutically acceptable organic primary, secondary or tertiaryamine. Representative pharmaceutically acceptable cations include alkalior alkaline earth salts such as lithium, sodium, potassium, calcium,magnesium, and aluminum salts and the like. Illustrative examples ofsome of the bases that can be used include sodium hydroxide, potassiumhydroxide, choline hydroxide, sodium carbonate, N⁺(C₁₋₄ alkyl)₄, and thelike. Representative organic amines useful for the formation of baseaddition salts include ethylamine, diethylamine, ethylenediamine,ethanolamine, diethanolamine, piperazine and the like. This inventionalso envisions the quaternization of any basic nitrogen-containinggroups of the compounds disclosed herein. Water or oil-soluble ordispersible products may be obtained by such quaternization. See, forexample, Berge et al., supra.

It should be understood that a reference to a salt includes the solventaddition forms or crystal forms thereof, particularly solvates orpolymorphs. Solvates contain either stoichiometric or non-stoichiometricamounts of a solvent, and are often formed during the process ofcrystallization with pharmaceutically acceptable solvents such as water,ethanol, and the like. Hydrates are formed when the solvent is water, oralcoholates are formed when the solvent is alcohol. Polymorphs includethe different crystal packing arrangements of the same elementalcomposition of a compound. Polymorphs usually have different X-raydiffraction patterns, infrared spectra, melting points, density,hardness, crystal shape, optical and electrical properties, stability,and solubility. Various factors such as the recrystallization solvent,rate of crystallization, and storage temperature may cause a singlecrystal form to dominate.

Pharmaceutically acceptable prodrugs of the compounds of this inventioninclude, but are not limited to, esters, carbonates, thiocarbonates,N-acyl derivatives, N-acyloxyalkyl derivatives, quaternary derivativesof tertiary amines, N-Mannich bases, Schiff bases, aminoacid conjugates,phosphate esters, metal salts and sulfonate esters.

Suitable positions for derivatization of the compounds of the inventionto create “prodrugs” include but are not limited, to, 2-aminosubstitution. Those of ordinary skill in the art have the knowledge andmeans to accomplish this without undue experimentation. Various forms ofprodrugs are well known in the art. See for example Design of Prodrugs,Bundgaard, A. Ed., Elseview, 1985 and Method in Enzymology, Widder, K.et al., Ed.; Academic, 1985, vol. 42, p. 309-396; Bundgaard, H. “Designand Application of Prodrugs” in A Textbook of Drug Design andDevelopment, Krosgaard-Larsen and H. Bundgaard, Ed., 1991, Chapter 5, p.113-191; and Bundgaard, H., Advanced Drug Delivery Review, 1992, 8,1-38, each of which is incorporated herein by reference.

The term “prodrugs” as employed herein includes, but is not limited to,the following groups and combinations of these groups, for example biscarbomates, or a carbomate and an acyloxyalkyl ester or a carbomate andan amide, etc.

Amine prodrugs;

Hydroxy prodrugs include, but are not limited to acyloxyalkyl esters,alkoxycarbonyloxyalkyl esters, alkyl esters, aryl esters and disulfidecontaining esters.

The term “alkyl” as used herein, alone or in combination, refers to anoptionally substituted straight-chain, or optionally substitutedbranched-chain saturated hydrocarbon monoradical having from one toabout thirty carbons, more preferably one to twelve carbons. Examples ofalkyl radicals include methyl, ethyl, n-propyl, isopropyl, n-butyl,isobutyl, sec-butyl, tert-butyl, tert-amyl, pentyl, hexyl, heptyl, octyland the like.

The term “cycloalkyl” as used herein, alone or in combination, refers tocyclic alkyl monoradicals which include monocyclic, bicyclic, tricyclic,and higher multicyclic alkyl radicals wherein each cyclic moiety hasfrom three to about eight carbon atoms. Examples of cycloalkyl radicalsinclude cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl and the like.

The term “lower alkyl” as used herein, alone or in combination, refersto an alkyl containing fewer carbon atoms, e.g., one containing from oneto about six carbon atoms.

The term “alkenyl” as used herein, alone or in combination, refers to anoptionally substituted straight-chain, or optionally substitutedbranched-chain hydrocarbon radical having one or more carbon-carbondouble-bonds and having from two to about thirty carbon atoms, morepreferably two to about eighteen carbons. Examples of alkenyl radicalsinclude ethenyl, propenyl, butenyl, 1,3-butadienyl and the like.

The term “cycloalkenyl” as used herein, alone or in combination, refersto to cyclic alkenyl radicals which include monocyclic, bicyclic,tricyclic, and higher multicyclic alkenyl radicals wherein each cyclicmoiety has from three to about eight carbon atoms.

The term “lower alkenyl” as used herein, alone or in combination, refersto an alkenyl having from two to about six carbons.

The term “alkynyl” as used herein, alone or in combination, refers to anoptionally substituted straight-chain or optionally substitutedbranched-chain hydrocarbon radical having one or more carbon-carbontriple-bonds and having from two to about thirty carbon atoms, morepreferably from two to about twelve carbon atoms, or from two to aboutsix carbon atoms, as well as those having from two to about four carbonatoms. Examples, of alkynyl radicals include ethynyl, 2-propynyl,2-butynyl, 1,3-butadiynyl and the like.

The term “cycloalkynyl” as used herein, alone or in combination, refersto cyclic alkynyl radicals that include monocyclic, bicyclic, tricyclic,and higher multicyclic alkynyl radicals wherein each cyclic moiety hasfrom three to about eight carbon atoms.

The term “lower alkynyl” as used herein, alone or in combination, refersto an alkynyl having from two to about six carbons.

The terms “heteroalkyl”, “heteroalkenyl” and “heteroalklynyl” as usedherein, alone or in combination, refer to include optionally substitutedalkyl, alkenyl and alkynyl structures, as described above, and whichhave one or more skeletal chain atoms selected from an atom other thancarbon, e.g., oxygen, nitrogen, sulfur, phosphorous or combinationsthereof.

The terms “lower heteroalkyl”, “lower heteroalkenyl” and “lowerheteroalkynyl” as used herein, alone or in combination, refer to,respectively, a heteroalkyl, heteroalkenyl and heteroalkynyl having fromtwo to about six carbons.

The term “alkylene” as used herein, alone or in combination, refers to adivalent hydrocarbyl group; because it is divalent, it can link twoother groups together. Typically it refers to —(CH₂)_(n)— where n is 1-8and preferably n is 14, though where specified, an alkylene can also besubstituted by other groups, and can be of other lengths, and the openvalences need not be at opposite ends of a chain. Thus —CH(Me)- and—C(Me)₂- may also be referred to as alkylenes, as can a cyclic groupsuch as cyclopropan-1,1-diyl. Where an alkylene group is substituted,the substituents include those typically present on alkyl groups asdescribed herein.

The term “lower alkylene” as used herein, alone or in combination,refers to an alkylene group containing fewer carbon atoms, e.g., onecontaining from one to about six carbon atoms.

The term “carbon chain” as used herein, alone or in combination, refersto any alkyl, alkenyl, alkynyl, or heteroalkyl, heteroalkenyl, orheteroalkynyl group, which is linear, cyclic, or any combinationthereof. If the chain is part of a linker and that linker comprises oneor more rings as part of the core backbone, for purposes of calculatingchain length, the “chain” only includes those carbon atoms that composethe bottom or top of a given ring and not both, and where the top andbottom of the ring(s) are not equivalent in length, the shorter distanceshall be used in determining the chain length. If the chain containsheteroatoms as part of the backbone, those atoms are not calculated aspart of the carbon chain length.

The term “membered ring” as used herein, alone or in combination, refersto any cyclic structure, including aromatic, heteroaromatic, alicyclic,heterocyclic, monocyclic, polycyclic, and fused rings. The term“membered” is meant to denote the number of skeletal (or ring) atomsthat constitute the ring system. Thus, for example, pyrrole,pyrrolidine, succinimide, maleimide, tetrahydrofuran and thiophene arefive-membered rings; pyridine, pyran, morpholine, piperazine, piperidineand pyrimidine are six-membered rings; and phthalimide, indole andindane are nine membered fused rings.

The term “aryl” as used herein, alone or in combination, refers to anoptionally substituted aromatic hydrocarbon monoradical of six to abouttwenty ring atoms, and includes mono-aromatic rings and fused aromaticrings. A fused aromatic ring radical contains from two to four fusedrings where the ring of attachment is an aromatic ring, and the otherindividual rings within the fused ring may be aromatic, heteroaromatic,alicyclic or heterocyclic. Further, the term aryl includes mono-aromaticrings and fused aromatic rings containing from six to about twelvecarbon atoms, as well as those containing from six to about ten carbonatoms. Examples of aryl groups include, without limitation, phenyl,naphthyl, anthryl, chrysenyl, and benzopyrenyl ring systems.

The term “lower aryl” as used herein, alone or in combination, refers toan aryl having six to about ten skeletal ring carbons, e.g., phenyl andnaphthyl ring systems.

The term “heteroaryl” as used herein, alone or in combination, refers tooptionally substituted aromatic radicals containing from about five toabout twenty skeletal ring atoms and where one or more of the ring atomsis a heteroatom such as, for example, oxygen, nitrogen, sulfur, seleniumor phosphorus. The term heteroaryl includes optionally substitutedmono-heteroaryl radicals and fused heteroaryl radicals having at leastone heteroatom (e.g., quinoline, benzothiazole). A fused heteroarylradical may contain from two to four fused rings where the ring ofattachment is a heteroaromatic ring, the other individual rings withinthe fused ring system may be aromatic, heteroaromatic, alicyclic orheterocyclic. The term heteroaryl also includes mono-heteroaryls orfused heteroaryls having from five to about twelve skeletal ring atoms,as well as those having from five to about ten skeletal ring atoms.Examples of heteroaryls include, without limitation, furanyl,benzofuranyl, chromenyl, pyridyl, pyrrolyl, indolyl, quinolinyl,pyridyl-N-oxide, pyrimidyl, pyrazinyl, imidazolyl, pyrazolyl, oxazolyl,isoxazolyl, benzothiozolyl, benzimidazolyl, benzoxazolyl,benzothiadiazolyl, benzoxadiazolyl, benzotriazolyl, quinolinyl,isoquinolinyl, indolyl, purinyl, indolizinyl, thienyl and the like andtheir oxides.

The term “lower heteroaryl” as used herein, alone or in combination,refers to a heteroaryl having five to about ten skeletal ring atoms,e.g., pyridyl, thienyl, pyrimidyl, pyrazinyl, pyrrolyl, or furanyl.

The terms “alicyclic” and “alicyclyl” as used herein, alone or incombination, refer to an optionally substituted saturated or unsaturatednonaromatic hydrocarbon ring system containing from three to abouttwenty ring atoms. The term alicyclic includes mono-alicyclic and fusedalicyclic radicals. A fused alicyclic may contain from two to four fusedrings where the ring of attachment is an alicyclic ring, and the otherindividual rings within the fused-alicyclic radical may be aromatic,heteroaromatic, alicyclic and heterocyclic. The term alicyclic alsoincludes mono-alicyclic and fused alicyclic radicals containing fromthree to about twelve carbon atoms, as well as those containing fromthree to about ten carbon atoms. Examples of alicyclics include, withoutlimitation, cyclopropyl, cyclopropenyl, cyclobutyl, cyclopentyl,cyclodecyl, cyclododecyl, cyclopentadienyl, indanyl, andcyclooctatetraenyl ring systems.

The terms “lower alicyclic” and “lower alicyclyl” as used herein, aloneor in combination, refer to an alicyclic having three to about tenskeletal ring carbons, e.g., cyclopropyl, cyclopropenyl, cyclobutyl,cyclopentyl, decalinyl, and cyclohexyl.

The terms “heterocyclic” and “heterocyclyl” as used herein, alone or incombination, refer to optionally substituted saturated or unsaturatednonaromatic ring radicals containing from five to about twenty ringatoms where one or more of the ring atoms are heteroatoms such as, forexample, oxygen, nitrogen, sulfur, and phosphorus. The term heterocyclicincludes mono-heterocyclic and fused heterocyclic ring radicals. A fusedheterocyclic radical may contain from two to four fused rings where theattaching ring is a heterocyclic, and the other individual rings withinthe fused heterocyclic radical may be aromatic, heteroaromatic,alicyclic or heterocyclic. The term heterocyclic also includesmono-heterocyclic and fused alicyclic radicals having from five to abouttwelve skeletal ring atoms, as well as those having from five to aboutten skeletal ring atoms. Example of heterocyclics include withoutlimitation, tetrahydrofuranyl, benzodiazepinyl, tetrahydroindazolyl,dihyroquinolinyl, and the like.

The terms “lower heterocyclic” and “lower heterocyclyl” as used herein,alone or in combination, refer to a heterocyclic ring system having fiveto about ten skeletal ring atoms, e.g., dihydropyranyl, pyrrolidinyl,dioxolanyl, piperidinyl, piperazinyl, and the like.

The term “alkylaryl” as used herein, alone or in combination, refers toan aryl radical as defined above in which at least one H atom isreplaced by an alkyl radical as defined above, such as, for example,tolyl, xylyl and the like.

The terms “arylalkyl” and “araalkyl” as used herein, alone or incombination, refer to an alkyl radical as defined above in which atleast one H atom is replaced by an aryl radical as defined above, suchas, for example, benzyl, 2-phenylethyl and the like.

The term “heteroarylalkyl” as used herein, alone or in combination,refers to an alkyl radical as defined above in which at least one H atomis replaced by a heteroaryl radical as defined above, each of which maybe optionally substituted.

The term “alkoxy” as used herein, alone or in combination, refers to analkyl ether radical, alkyl-O—, wherein the term alkyl is defined asabove. Examples of alkoxy radicals include methoxy, ethoxy, n-propoxy,isopropoxy, n-butoxy, iso-butoxy, sec-butoxy, tert-butoxy and the like.

The term “lower alkoxy” as used herein, alone or in combination, refersto an alkoxy group having one to about six carbon atoms.

The term “aryloxy” as used herein, alone or in combination, refers to anaryl ether radical wherein the term aryl is defined as above. Examplesof aryloxy radicals include phenoxy, thienyloxy and the like.

The terms “alkylthio” and “thioalkyl” as used herein, alone or incombination, refer to an alkyl thio radical, alkyl-S—, wherein the termalkyl is as defined above.

The term “arylthio” as used herein, alone or in combination, refers toan aryl thio radical, aryl-S—, wherein the term aryl is as definedabove.

The term “heteroarylthio” as used herein, alone or in combination,refers to the group heteroaryl-S—, wherein the term heteroaryl is asdefined above.

The term “acyl” as used herein, alone or in combination, refers to aradical —C(O)R where R includes alkyl, alkenyl, alkynyl, aryl,heteroaryl, alicyclic, heterocyclic, arylalkyl or heteroarylalkylwherein the alkyl, alkenyl, alkynyl, aryl, heteroaryl, alicyclic,heterocyclic, arylalkyl or heteroaryl alkyl groups may be optionallysubstituted.

The term “acyloxy” as used herein, alone or in combination, refers tothe ester group —OC(O)R, where R is H, alkyl, alkenyl, alkynyl, aryl,heteroaryl, alicyclic, heterocyclic, arylalkyl, or heteroarylalkylwherein the alkyl, alkenyl, alkynyl, aryl, heteroaryl, alicyclic,heterocyclic, arylalkyl or heteroarylalkyl may be optionallysubstituted.

The term “carboxy esters” as used herein, alone or in combination,refers to —C(O)OR where R is alkyl, aryl or arylalkyl, wherein thealkyl, aryl and arylalkyl groups may be optionally substituted. The term“BOC” as used herein, alone or in combination, refers to —C(O)Otbutyl.The term “carboxarido” as used herein, alone or in combination, refersto

where each of R and R′ are independently selected from the groupconsisting of H, alkyl, aryl, heteroaryl, alicyclic, heterocyclic,arylalkyl and heteroarylalkyl, wherein the alkyl, aryl, heteroaryl,alicyclic, heterocyclic, or arylalkyl groups may be optionallysubstituted.

The terms “thioamide” and “thioamidyl” as used herein, alone or incombination, refer to

where each of R and R′ are independently selected from the groupconsisting of H, alkyl, aryl, heteroaryl, alicyclic, heterocyclic,arylalkyl and heteroarylalkyl, wherein the alkyl, aryl, heteroaryl,alicyclic, heterocyclic, or arylalkyl groups may be optionallysubstituted.

The term “oxo” as used herein, alone or in combination, refers to ═O.

The term “halogen” as used herein, alone or in combination, refers to F,Cl, Br and I.

The terms “haloalkyl”, “haloalkenyl”, “haloalkynyl” and “haloalkoxy” asused herein, alone or in combination, refer to alkyl, alkenyl, alkynyland alkoxy structures, as described above, that are substituted with oneor more fluorines, chlorines, bromrines or jodines, or with combinationsthereof.

The terms “perhaloalkyl”, “perhaloalkyloxy” and “perhaloacyl” as usedherein, alone or in combination, refer to alkyl, alkyloxy and acylradicals as described above, in which all the H atoms are replaced byfluorines, chlorines, bromines or iodines, or combinations thereof.

The terms “lower perhaloalkyl”, “lower perhaloalkyloxy” and “lowerperhaloacyl” as used herein, alone or in combination, refer toperhaloalkyl, perhaloalkyloxy and perhaloacyl radicals as describedabove, having from two to about six carbons.

The terms cycloalkyl, arylalkyl, aryl, heteroaryl, alicyclic,heterocyclic, alkyl, alkynyl, alkenyl, haloalkyl, and heteroalkylinclude optionally substituted cycloalkyl, arylalkyl, aryl, heteroaryl,alicyclic, heterocyclic, alkyl, alkynyl, alkenyl, haloalkyl andheteroalkyl groups.

The term “alkylsilyl” as used herein, alone or in combination, refers to—NRR′R″ where R, R′ and R″ are alkyls.

The term “lower alkylsilyl” as used herein, alone or in combination,refers to —NRR′R″ where R, R′ and R″ are lower alkyls.

The term “amino” as used herein, alone or in combination, refers to—NH₂.

The term “alkylamino” as used herein, alone or in combination, refers tothe group —NHR where R is alkyl.

The term “aminoalkyl” as used herein, alone or in combination, refers tothe group -alkylene-NH₂, wherein alkylene is as defined herein.

The term “dialkylamino” as used herein, alone or in combination, refersto the group —NRR′ where R and R′ are alkyls.

The term “arylalkylamino” as used herein, alone or in combination,refers to the group —NRR′ where R is alkyl, and R′ is aryl.

The term “diarylamino” as used herein, alone or in combination, refersto the group —NRR′ where R and R′ are aryls.

The term “heteroarylamino” as used herein, alone or in combination,refers to the group —NHR where R is heteroaryl.

The term “diheteroarylamino” as used herein, alone or in combination,refers to the group —NRR′ where R and R′ are heteroaryls.

The term “arylheteroarylamino” as used herein, alone or in combination,refers to the group —NRR′ where R is aryl, and R′ is heteroaryl.

The term “carbamyl” as used herein, alone or in combination, refers tothe —NHC(O)OR and —OC(O)NHR groups, wherein R may be, but is not limitedto alkyl, alkenyl, alkynyl, aryl, alicyclic, heterocyclic, aryl,heteroaryl, arylalkyl and heteroarylalkyl, wherein the alkyl, alkenyl,alkynyl, aryl, alicyclic, heterocyclic, aryl, heteroaryl, arlalkyl andheteroarylalkyl groups may be optionally substituted. Non-limitingexamples of sulfanyl groups include methylsulfanyl (—SCH₃) andiso-propylsulfanyl (—SCH(CH₃)₂) and the like.

The term “uryl” as used herein, alone or in combination, refers to the—NHC(O)NHR group, wherein R may be, but is not limited to alkyl,alkenyl, alkynyl, aryl, alicyclic, heterocyclic, aryl, heteroaryl,arylalkyl and heteroarylalkyl, wherein the alkyl, alkenyl, alkynyl,aryl, alicyclic, heterocyclic, aryl, heteroaryl, arylalkyl andheteroarylalkyl groups may be optionally substituted. Non-limitingexamples of sulfanyl groups include methylsulfanyl (—SCH₃) andiso-propylsulfanyl (—SCH(CH₃)₂) and the like.

The term “thiouryl” as used herein, alone or in combination, refers tothe —NHC(S)NHR group, wherein R may be, but is not limited to alkyl,alkenyl, alkynyl, aryl, alicyclic, heterocyclic, aryl, heteroaryl,arylalkyl and heteroarylalkyl, wherein the alkyl, alkenyl, alkynyl,aryl, alicyclic, heterocyclic, aryl, heteroaryl, arylalkyl andheteroarylalkyl groups may be optionally substituted. Non-limitingexamples of sulfanyl groups include methylsulfanyl (—SCH₃) andiso-propylsulfanyl (—SCH(CH₃)₂) and the like.

The term “guanidinyl” as used herein, alone or in combination, refers tothe —NHC(NH)NHR group, wherein R may be, but is not limited to alkyl,alkenyl, alkynyl, aryl, alicyclic, heterocyclic, aryl, heteroaryl,arylalkyl and heteroarylalkyl, wherein the alkyl, alkenyl, alkynyl,aryl, alicyclic, heterocyclic, aryl, heteroaryl, arylalkyl andheteroarylalkyl groups may be optionally substituted.

Non-limiting examples of sulfanyl groups include methylsulfanyl (—SCH₃)and iso-propylsulfanyl (—SCH(CH₃)₂) and the like.

The terms “sulfide” and “thioether” as used herein, alone or incombination, refer to a sulfur atom covalently linked to two atoms; theformal oxidation state of said sulfur is (II). These terms may be usedinterchangeably.

The term “sulfanyl” as used herein, alone or in combination, refers tothe —S—R group, wherein R may be, but is not limited to alkyl, alkenyl,alkynyl, aryl, alicyclic, heterocyclic, aryl, heteroaryl, arylalkyl andheteroarylalkyl, wherein the alkyl, alkenyl, alkynyl, aryl, alicyclic,heterocyclic, aryl, heteroaryl, arylalkyl and heteroarylalkyl groups maybe optionally substituted. Non-limiting examples of sulfanyl groupsinclude methylsulfanyl (—SCH₃) and iso-propylsulfanyl (—SCH(CH₃)₂) andthe like.

The term “sulfoxide” as used herein, alone or in combination, refers toa sulfur atom covalently linked to three atoms, at least one of which isan oxygen atom; the formal oxidation state of said sulfur atom is (IV).

The term “sulfinyl” as used herein, alone or in combination, refers tothe groups —S(O)—R, wherein R may be, but is not limited to alkyl,alkenyl, alkynyl, aryl, alicyclic, heterocyclic, aryl, heteroaryl,arylalkyl and heteroarylalkyl, wherein the alkyl, alkenyl, alkynyl,aryl, alicyclic, heterocyclic, aryl, heteroaryl, arylalkyl andheteroarylalkyl groups may be optionally substituted. A non-limitingexample of a sulfinyl group includes methylsulfinyl (—S(O)CH₃) and thelike. The term “sulfone” as used herein, alone or in combination, refersto a sulfur atom covalently linked to four atoms, at least two of whichare oxygen atoms; the formal oxidation state of said sulfur atom is(VI).

The term “sulfonyl” as used herein, alone or in combination, refers tothe groups —S(O₂)—R, wherein R may be, but is not limited to alkyl,alkenyl, alkynyl, aryl, alicyclic, heterocyclic, aryl, heteroaryl,arylalkyl and heteroarylalkyl, wherein the alkyl, alkenyl, alkynyl,aryl, alicyclic, heterocyclic, aryl, heteroaryl, arylalkyl andheteroarylalkyl groups may be optionally substituted. A non-limitingexample of a sulfonyl group includes methylsulfonyl (—S(O₂)CH₃) and thelike. The term “phosphite” as used herein, alone or in combination,refers to a phosphorus atom covalently linked to three carbon atoms,wherein the formal oxidation state of said phosphorus is (III).

The term “phosphinyl” as used herein, alone or in combination, refers tothe monoradical derived from a phosphite group, as defined above.

The term “phosphonate” as used herein, alone or in combination, refersto a phosphorus atom covalently linked to four atoms, three of which areoxygen and one of which is carbon wherein the formal oxidation state ofsaid phosphorus is (V).

The term “phosphonyl” as used herein, alone or in combination, refers tothe monoradical derived from a phosphonate group, as defined above.

The term “phosphate” as used herein, alone or in combination, refers toa phosphorus atom covalently linked to four oxygen atoms, wherein theformal oxidation state of said phosphorus is (V).

The term “phosphatidyl” as used herein, alone or in combination, refersto the monoradical derived from a phosphate group, as defined above.

The term “phosphoramide” as used herein, alone or in combination, refersto a phosphorus atom covalently linked to four atoms, three of which arenitrogen and one of which is oxygen wherein the formal oxidation stateof said phosphorus is (V).

The term “phosphoramidyl” as used herein, alone or in combination,refers to the monoradical derived from a phosphoramide group, as definedabove.

The terms “optional” or “optionally” mean that the subsequentlydescribed event or circumstance may but need not occur, and that thedescription includes instances where the event or circumstance occursand instances in which it does not. For example, “aryl optionally mono-or di-substituted with an alkyl” means that the alkyl may but need notbe present, or either one alkyl or two may be present, and thedescription includes situations where the aryl is substituted with oneor two alkyls and situations where the aryl is not substituted with analkyl.

“Optionally substituted” groups may be substituted or unsubstituted. Thesubstituents of an “optionally substituted” group may include, withoutlimitation, one or more substituents independently selected from thefollowing groups or designated subsets thereof: lower alkyl, loweralkenyl, lower alkynyl, lower aryl, heteroaryl, alicyclic, heterocyclic,arylalkyl, heteroarylalkyl, lower alkoxy, lower aryloxy, amino,alkylamino, dialkylamino, diarylalkylamino, alkylthio, arylthio,heteroarylthio, oxo, oxa, acyl (—C(O)R), (—C(O)), carboxyesters(—C(O)OR), carboxamido (—C(O)NH₂), carboxy, acyloxy, —H, halo, —CN,—NO₂, —N₃, —SH, —OH, —(O)CH₃, perhaloalkyl, perhaloalkoxy, perhaloacyl,guanidine, pyridinyl, thiophene, furanyl, indole, indazole, esters,amides, phosphonates, phosphonic acid, phosphates, phosphoramides,sulfonates, sulfones, sulfates, sulphonamides, carbamates, ureas,thioureas, thioamides and thioalkyls. An optionally substituted groupmay be unsubstituted (e.g., —CH₂CH₃), fully substituted (e.g., —CF₂CF₃),monosubstituted (e.g., —CH₂CH₂F) or substituted at a level anywherein-between fully substituted and monosubstituted (e.g., —CH₂CF₃).

The term “pyridine-1-oxy” also means “pyridine-N-oxy”.

The term “heteroaryl group is substituted with 2 to 5 substituents”encompasses 1-oxy-pyridyl or N-oxy-pyridyl having 1 to 4 substituents,i.e. the oxygen atom of the pyridine-N-oxide should be counted as asubstituent.

Some of the compounds of the present invention may contain one or morechiral centers and therefore may exist in enantiomeric anddiastereomeric forms. The scope of the present invention is intended tocover all isomers per se, as well as mixtures of cis and trans isomers,mixtures of diastereomers and racemic mixtures of enantiomers (opticalisomers) as well. Further, it is possible using well known techniques toseparate the various forms, and some embodiments of the invention mayfeature purified or enriched species of a given enantiomer ordiastereomer.

Some of the compounds of the present invention may exist in tautomericforms. The scope of the present invention is intended to cover alltautomers. As a non-limiting example, compounds of formula (I), whereinR⁰ is —SR⁸ or —OR⁸, and R⁸ is hydrogen, could exist as either of thetautomers below:

A “pharmacological composition” refers to a mixture of one or more ofthe compounds described herein, or pharmaceutically acceptable saltsthereof, with other chemical components, such as pharmaceuticallyacceptable carriers and/or excipients. The purpose of a pharmacologicalcomposition is to facilitate administration of a compound to anorganism.

The phrase “pharmaceutically acceptable carrier” as used herein means apharmaceutically-acceptable material, composition or vehicle, such as aliquid or solid filler, diluent, excipient, solvent or encapsulatingmaterial, involved in carrying or transporting the subject agent fromone organ, or portion of the body, to another organ, or portion of thebody. Each carrier must be “acceptable” in the sense of being compatiblewith the other ingredients of the formulation and not injurious to thepatient. Some examples of materials which can serve aspharmaceutically-acceptable carriers include: (1) sugars, such aslactose, glucose and sucrose; (2) starches, such as corn starch andpotato starch; (3) cellulose, and its derivatives, such as sodiumcarboxymethyl cellulose, ethyl cellulose and cellulose acetate; (4)powdered tragacanth; (5) malt; (6) gelatin; (7) talc; (8) excipients,such as cocoa butter and suppository waxes; (9) oils, such as peanutoil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil andsoybean oil; (10) glycols, such as propylene glycol; (11) polyols, suchas glycerin, sorbitol, mannitol and polyethylene glycol; (12) esters,such as ethyl oleate and ethyl laurate; (13) agar; (14) bufferingagents, such as magnesium hydroxide and aluminum hydroxide; (15) alginicacid; (16) pyrogen-free water; (17) isotonic saline; (18) Ringer'ssolution; (19) ethyl alcohol; (20) phosphate buffer solutions; and (21)other non-toxic compatible substances employed in pharmaceuticalformulations. A physiologically acceptable carrier should not causesignificant irritation to an organism and does not abrogate thebiological activity and properties of the administered compound.

An “excipient” refers to an inert substance added to a pharmacologicalcomposition to further facilitate administration of a compound. Examplesof excipients include but are not limited to calcium carbonate, calciumphosphate, various sugars and types of starch, cellulose derivatives,gelatin, vegetable oils and polyethylene glycols.

A “pharmaceutically effective amount” means an amount which is capableof providing a therapeutic and/or prophylactic effect. The specific doseof compound administered according to this invention to obtaintherapeutic and/or prophylactic effect will, of course, be determined bythe particular circumstances surrounding the case, including, forexample, the specific compound administered, the route ofadministration, the condition being treated, and the individual beingtreated. A typical daily dose (administered in single or divided doses)will contain a dosage level of from about 0.01 mg/kg to about 50-100mg/kg of body weight of an active compound of the invention. Preferreddaily doses generally will be from about 0.05 mg/kg to about 20 mg/kgand ideally from about 0.1 mg/kg to about 10 mg/kg. Factors such asclearance rate, half-life and maximum tolerated dose (MTD) have yet tobe determined but one of ordinary skill in the art can determine theseusing standard procedures.

In some method embodiments, the preferred therapeutic effect is theinhibition, to some extent, of the growth of cells characteristic of aproliferative disorder, e.g., breast cancer. A therapeutic effect willalso normally, but need not, relieve to some extent one or more of thesymptoms other than cell growth or size of cell mass. A therapeuticeffect may include, for example, one or more of 1) a reduction in thenumber of cells; 2) a reduction in cell size; 3) inhibition (i.e.,slowing to some extent, preferably stopping) of cell infiltration intoperipheral organs, e.g., in the instance of cancer metastasis; 3)inhibition (i.e., slowing to some extent, preferably stopping) of tumormetastasis; 4) inhibition, to some extent, of cell growth; and/or 5)relieving to some extent one or more of the symptoms associated with thedisorder. As used herein, the term IC₅₀ refers to an amount,concentration or dosage of a particular test compound that achieves a50% inhibition of a maximal response in an assay that measures suchresponse. In some method embodiments of the invention, the “IC₅₀” valueof a compound of the invention can be greater for normal cells than forcells exhibiting a proliferative disorder, e.g., breast cancer cells.The value depends on the assay used.

By a “standard” is meant a positive or negative control. A negativecontrol in the context of Her2 expression levels is, e.g., a samplepossessing an amount of Her2 protein that correlates with a normal cell.A negative control may also include a sample that contains no Her2protein. By contrast, a positive control does contain Her2 protein,preferably of an amount that correlates with overexpression as found inproliferative disorders, e.g., breast cancers. The controls may be fromcell or tissue samples, or else contain purified ligand (or absentligand), immobilized or otherwise. In some embodiments, one or more ofthe controls may be in the form of a diagnostic “dipstick.”

By “selectively targeting” is meant affecting one type of cell to agreater extent than another, e.g., in the case of cells with high asopposed to relatively low or normal Her2 levels.

Compounds of the Invention

Compounds of the invention are related to alkynylpyrrolo[2,3-d]pyrimidines of Formula I

and their polymorphs, solvates, esters, tautomers, diastereomers,enantiomers, pharmaceutically acceptable salts or prodrugs thereof,which show utility for inhibiting HSP90 and treating and preventingdiseases that are HSP90-dependent, wherein:

R⁰ is selected from the group consisting of hydrogen, halogen, loweralkyl, —CN, —SR⁸, —OR⁸, and —NHR⁸;

R¹ is selected from the group consisting of halogen, —OR¹¹, —SR¹¹ andlower alkyl;

R² is —NHR⁸;

R³ is selected from the group consisting of hydrogen, —CN, —C(O)OH,—OR¹¹, —SR¹¹, —C(O)R⁹, —NR⁵R¹⁰, lower alkyl, lower alkenyl, loweralkynyl, lower perhaloalkyl, lower alkylsilyl, aryl, heteroaryl,alicyclyl and heterocyclyl, all optionally substituted, wherein:

-   -   the aryl, heteroaryl, alicyclyl and heterocyclyl groups are        mono-, bi- or tri-cyclic;    -   R⁸ and R¹⁰ taken together with the N atom to which they are        attached optionally form an optionally substituted ring        comprising 3-7 ring atoms, wherein, in addition to said N atom,        0-3 of the ring atoms are heteroatoms selected from the group        consisting of O, S and N;    -   the optional substituents on R³ are selected from the group        consisting of lower alkyl, lower alkenyl, lower alkynyl, —CN,        —C(O)OH, —NO₂, —SR⁸, —OR⁸, —C(O)R⁹, —NR⁸R⁸, lower aryl,        heteroaryl, alicyclyl, lower heterocyclyl, arylalkyl,        heteroarylalkyl, amino, alkylamino, dialkylamino,        arylalkylamino, diarylamino, heteroarylamino, diheteroarylamino,        arylheteroarylamino, oxo, perhaloalkyl, perhaloalkoxy,        perhaloacyl, guanidinyl, pyridinyl, thiophenyl, furanyl,        indolyl, indazolyl, phosphonyl, phosphatidyl, phosphoramidyl,        sulfanyl, sulfinyl, sulfonyl, sulphonamidyl, carbamyl, uryl,        thiouryl and thioamidyl, wherein    -   R⁸ and R⁸ taken together with the N atom to which they are        attached optionally form an optionally substituted ring        comprising 3-7 ring atoms, wherein, in addition to said N atom,        0-3 of the ring atoms are heteroatoms selected from the group        consisting of O, S and N;

R⁴ is selected from the group consisting of optionally substituted loweralkylene, —C(R¹²)₂—, —C(O)—, —C(S)—, —S(O)— and —SO₂—;

R⁵ is selected from the group consisting of aryl, heteroaryl, alicyclyland heterocyclyl, wherein:

-   -   the aryl group is substituted with 2 to 5 substituents;    -   the heteroaryl group is substituted with 2 to 5 substituents;    -   the alicyclyl group is substituted with 3 to 5 substituents;    -   the heterocyclyl group is substituted with 3 to 5 substituents;    -   the substituents on R⁵ are selected from the group consisting of        halogen, lower alkyl, lower alkenyl, lower alkynyl, —CN,        —C(O)OH, —NO₂, —SR⁸, —OR⁸, —C(O)R⁹, —NR⁸R¹⁰, lower aryl, lower        heteroaryl, lower alicyclyl, lower heterocyclyl, arylalkyl,        heteroarylalkyl, thioalkyl, amino, alkylamino, dialkylamino,        arylalkylamino, oxo, perhaloalkyl, perhaloalkoxy, perhaloacyl,        guanidinyl, pyridinyl, thiophenyl, furanyl, indolyl, indazolyl,        phosphonyl, phosphatidyl, phosphoramidyl, sulfanyl, sulfinyl,        sulfonyl, sulphonamidyl, carbamyl, uryl, thiouryl and        thioamidyl, wherein    -   R⁸ and R¹⁰ taken together with the N atom to which they are        attached optionally form an optionally substituted ring        comprising 3-7 ring atoms, wherein, in addition to said N atom,        0-3 of the ring atoms are heteroatoms selected from the group        consisting of O, S and N;

R⁸ is selected from the group consisting of hydrogen, lower alkyl, loweralkenyl, lower alkynyl, lower heteroalkyl, lower heteroalkenyl, lowerheteroalkynyl, lower aryl, lower heteroaryl and —C(O)R⁹;

R⁹ is selected from the group consisting of H, lower alkyl, loweralkenyl, lower alkynyl, lower aryl, lower heteroaryl, —NR¹⁰R¹⁰ and—OR¹¹, wherein

R¹⁰ and R¹⁰ taken together with the N atom to which they are attachedoptionally form an optionally substituted ring comprising 3-7 ringatoms, wherein, in addition to said N atom, 0-3 of the ring atoms areheteroatoms selected from the group consisting of O, S and N;

R¹⁰ is selected from the group consisting of hydrogen, lower alkyl,lower alkenyl, lower alkynyl, lower heteroalkyl, lower heteroalkenyl,lower heteroalkynyl, lower aryl, lower heteroaryl and —C(O)R¹¹;

R¹¹ is selected from the group consisting of lower alkyl, lower alkenyl,lower alkynyl, lower aryl and lower heteroaryl; and

R¹² is selected from the group consisting of hydrogen and lower alkyl.

In some embodiments, R⁰ is hydrogen, halogen or —CN. In otherembodiments, R is hydrogen, lower alkyl, —SR⁸ or —OR⁸. In otherembodiments, R⁰ is hydrogen, —SR, —OR⁸ or —NHR⁸. In other embodiments,R⁰ is —SR⁸ or —OR⁸. In other embodiments, R⁰ is hydrogen.

In some embodiments, R⁰ is halogen or lower alkyl. In other embodiments,R¹ is —OR¹¹ or —SR¹¹. In some embodiments, R¹ is halogen. In someembodiments, R¹ is chloro or bromo. In some embodiments, R¹ is chloro.In some embodiments, R¹ is bromo.

In some embodiments, R² is —NH₂ or —NHC(O)R⁹. In other embodiments, R²is —NH-lower alkyl, —NH-lower alkenyl, —NH-lower alkynyl, —NH-lower arylor —NH-lower heteroaryl. In other embodiments, R² is —NHC(O)R⁹ where R⁹is lower alkyl, lower alkenyl, lower alkynyl, lower aryl or lowerheteroaryl. In other embodiments, R² is —NH₂. In other embodiments, R²is —NH(O)tBu.

In some embodiments, R³ is hydrogen, lower alkyl, lower alkenyl, loweralkynyl, lower perhaloalkyl, aryl, heteroaryl, alicyclyl, heterocyclyl,—CN or —C(O)R⁹, all optionally substituted. In other embodiments, R³ ishydrogen, lower alkyl, aryl, heteroaryl, alicyclyl, heterocyclyl or—C(O)R⁹, all optionally substituted. In other embodiments, R³ ishydrogen, lower alkyl, aryl, heteroaryl, or —C(O)R⁹, all optionallysubstituted. In other embodiments, R³ is substituted lower alkyl whereinthe substituent on the lower alkyl is selected from the group consistingof lower alkyl, —OR⁸, —C(O)R⁹ and —NR⁸R⁸. In other embodiments, R³ islower alkyl, aryl, heteroaryl, —CN or —C(O)R⁹, all optionallysubstituted. In other embodiments, R³ is hydrogen. In other embodiments,R³ is optionally substituted lower alkyl. In other embodiments, R³ isoptionally substituted phenyl or pyridinyl. In other embodiments, R³ is—(CH₂)_(n)OH, where n=1-3. In other embodiments, R³ is—(CH₂)_(m)C(R¹²)₂(CH₂)_(n)OH, where m=0-2 and n=1-2. In otherembodiments, R³ is —(CH₂)_(n)NR⁸R⁸, wherein n=1-3, and each R⁸ isindependently hydrogen or lower alkyl, or —NR⁸R⁸ are taken togetherforming an optionally substituted phthalimide or morpholin. In otherembodiments, R³ is —(CH₂)_(n)C(O)NR¹⁰R¹⁰, wherein n=1-3, and each R¹⁰ isindependently hydrogen or —C(O)R¹¹, or —NR¹⁰R¹⁰ are taken togetherforming an optionally substituted piperazine. In other embodiments, R³is optionally substituted lower alkylsilyl. In other embodiments, R³ is—CO₂Et. In other embodiments, R³ is —(CH₂)_(n)C(O)NR¹⁰R¹⁰ where n=1-3,and each R¹⁰ is independently hydrogen, or —C(O)R¹¹. In otherembodiments, R³ is —(CH₂)_(n)C(O)NHC(O)OtBu.

In some embodiments, R⁴ is optionally substituted lower alkylene,—C(O)—, —S(O)— or —SO₂—. In other embodiments, R⁴ is —CH₂—, —S(O)— or—SO₂. In other embodiments, R⁴ is —CH₂—. In other embodiments, R⁴ is—CH₂—.

In some embodiments, the aryl, heteroaryl, alicyclyl or heterocyclylgroup of R⁵ is monocyclic or bicyclic. In other embodiments, R⁵ issubstituted aryl or heteroaryl and the substituents on said aryl orheteroaryl are selected from the group consisting of halogen, loweralkoxy, lower alkyl, thioalkyl, amino, alkylamino, dialkylamino. Inother embodiments, R⁵ is substituted aryl or heteroaryl and thesubstituents on the aryl or heteroaryl are selected from the groupconsisting of halogen, lower alkoxy and lower alkyl.

In some embodiments, R⁰ is hydrogen, halogen, —SH, —OH, or —CN; R¹ ishalogen; and R² is —NH₂ or —NH—C(O)R.

In some embodiments, R¹ is chloro or bromo; R² is —NH₂ or —NH—C(O)R⁹;and R³ is lower alkyl, lower alkenyl, lower alkynyl, lower perhaloalkyl,lower aryl, or lower heteroaryl, all optionally substituted with —OR⁸,—NR⁸R⁸ or —C(O)R⁹.

In some embodiments, Ro is hydrogen, halogen or —CN; R² is —NH₂ or—NH—C(O)R⁹, and R⁴ is —CH₂—.

In other embodiments, R⁰ is hydrogen, halogen, —SH, —OH or —CN; R¹ ishalogen; R² is —NH₂; R³ is hydrogen, —OR¹¹, —SR¹¹, —NR⁸R⁸, lower alkyl,lower alkenyl, lower alkynyl, lower perhaloalkyl, lower aryl, or lowerheteroaryl, wherein the R⁸ in R³ is hydrogen, lower alkyl, lowerheteroalkyl, lower aryl, or —C(O)R⁹; R⁴ is —CH₂—; and R⁵ is aryl orheteroaryl, substituted with 2 to 5 substituents.

In some embodiments, R⁰ is selected from hydrogen, halogen and —CN; R¹is halogen; R² is NHR⁸; R³ is selected from the group consisting ofhydrogen, lower alkyl, lower alkenyl, lower alkynyl, lower perhaloalkyl,aryl, heteroaryl, alicyclyl and heterocyclyl, all optionallysubstituted;

R⁴ is —CHR¹²—; and R⁵ optionally substituted is aryl or heteroaryl.

In other embodiments, R⁰ is selected from hydrogen, halogen and —CN; R¹is halogen; R² is —NH₂; R³ is selected from the group consisting oflower alkyl, lower alkenyl, lower alkynyl, all optionally substitutedwith —OR⁸, —NR⁸R⁸ or —C(O)R⁹. R⁴ is —CH₂; and R⁵ is aryl or heteroaryl.

In other embodiments, R¹ is chloro or bromo; R² is —NH₂; and R⁵ is aphenyl having at least three substituents.

In other embodiments, R¹ is chloro or bromo, R² is —NH₂; and R⁵ is apyridyl having at least two substituents.

In other embodiments, R¹ is chloro or bromo; R² is —NH₂; and R⁵ is1-oxy-pyridyl (N-oxy-pyridyl) having at least two substituents.

In other selected embodiments, R⁰ is hydrogen; R¹ is chloro or bromo; R²is —NH₂ or —NHC(O)_(t)Bu; R³ is selected from the group consisting ofhydrogen, lower alkyl, aryl, heteroaryl, —CN, optionally substitutedlower alkylsilyl, —CO₂Et, —(CH₂)_(m)C(CH₃)_(n)OH where m=0-2 and n=1-2,—(CH₂)_(n)OH where n=1-3, and —(CH₂)_(n)NR⁸R¹⁰ wherein n=1-3, R⁵ and R¹⁰are independently hydrogen, lower alkyl, or are taken together forming apiperazine, a phthalimide or a morpholine;

R⁴ is —CH₂—; R⁵ is substituted aryl or heteroaryl, and the substituentson the aryl or heteroaryl are selected from the group consisting ofhalogen, lower alkoxy, lower alkyl, thioalkyl, amino, alkylamino anddialkylamino.

In some selected embodiments, R⁰ is hydrogen; R¹ is chloro or bromo; R²is —NH₂;

R³ is selected from the group consisting of hydrogen, lower alkyl,—(CH₂)_(n)OH, where n=1-3, —(CH₂)_(m)C(CH₃)_(n)OH, where m=0-2 andn=1-2, —(CH₂)₂C(O)NH₂, —(CH₂)OC(O)CH₂N(CH₃)₂, —(CH₂)_(n)NHC(O)OtBu and—(CH₂)_(n)NR⁸R⁸, wherein n=1-3, R⁸ and R⁸ are independently hydrogen,lower alkyl, or are taken together forming an optionally substitutedphthalimide or morpholine; R⁴ is —CH₂—; R⁵ is substituted heteroaryl,and the substituents on said heteroaryl are independently selected fromthe group consisting of lower alkoxy and lower alkyl.

In some selected embodiments, R⁰ is hydrogen; R¹ is chloro or bromo; R²is —NH₂;

R³ is selected from the group consisting of hydrogen, lower alkyl,—(CH₂)_(n)OH, where n=1-3, —(CH₂)_(m)C(CH₃)_(n)OH, where m=0-2 andn=1-2, —(CH₂)_(n)C(O)NR¹⁰R¹⁰ where n=1-3, and, R¹⁰ and R¹⁰ areindependently hydrogen or —C(O)R¹¹, or are taken together forming anoptionally substituted piperazine; R⁴ is —CH₂—; R⁵ is substitutedheteroaryl, and the substituents on said heteroaryl are independentlyselected from the group consisting of lower alkoxy and lower alkyl.

In some selected embodiments R³ is substituted lower alkyl, and in somefurther selected embodiments R³ is substituted lower alkyl, and thesubstituent on said lower alkyl is phosphonyl or phosphatidyl.

In other preferred embodiments, are the following compounds

It should be understood that any of the above described embodiments ofthe invention can be combined in anyway where practical; those ofordinary skill in the art will appreciate the ways the variousembodiments may be combined usefully within the spirit of the invention.A non-limiting list of compounds based on Formula I of the invention isexemplified in TABLE 1.

TABLE 1 Exemplary Compounds based on Formula I (I)

wherein R⁰ is H, and R⁴ is —CH₂—. No. Ex R¹ R² R³ R⁵ 1 Cl NH₂ H3,4,5-Trimethoxyphenyl 2 Cl NH₂ H 2-Chloro-3,4,5-trimethoxyphenyl 3 ClNH₂ H 2-Bromo-3,4,5-trimethoxyphenyl 4 Cl NH₂ H2-Iodo-3,4,5-trimethoxyphenyl 5 Cl NH₂ H 2-Fluoro-3,4,5-trimethoxyphenyl6 Cl NH₂ H 3,4,5-Trimethylphenyl 7 Cl NH₂ H2-Chloro-3,4,5-trimethylphenyl 8 Cl NH₂ H 2-Bromo-3,4,5-trimethylphenyl9 Cl NH₂ H 2-Iodo-3,4,5-trimethylphenyl 10 Cl NH₂ H2-Fluoro-3,4,5-trimethylphenyl 11 Cl NH₂ H 3,5-Dimethoxy-4-methylphenyl12 Cl NH₂ H 2-Chloro-3,5-dimethoxy-4-methylphenyl 13 Cl NH₂ H2-Bromo-3,5-dimethoxy-4-methylphenyl 14 Cl NH₂ H2-Iodo-3,5-dimethoxy-4-methylphenyl 15 Cl NH₂ H2-Fluoro-3,5-dimethoxy-4-methylphenyl 16 Cl NH₂ 2-Py3,4,5-Trimethoxyphenyl 17 Cl NH₂ 2-Py 2-Chloro-3,4,5-trimethoxyphenyl 18Cl NH₂ 2-Py 2-Bromo-3,4,5-trimethoxyphenyl 19 Cl NH₂ 2-Py2-Iodo-3,4,5-trimethoxyphenyl 20 Cl NH₂ 2-Py2-Fluoro-3,4,5-trimethoxyphenyl 21 Cl NH₂ 2-Py 3,4,5-Trimethylphenyl 22Cl NH₂ 2-Py 2-Chloro-3,4,5-trimethylphenyl 23 Cl NH₂ 2-Py2-Bromo-3,4,5-trimethylphenyl 24 Cl NH₂ 2-Py2-Iodo-3,4,5-trimethylphenyl 25 Cl NH₂ 2-Py2-Fluoro-3,4,5-trimethylphenyl 26 Cl NH₂ 2-Py3,5-Dimethoxy-4-methylphenyl 27 Cl NH₂ 2-Py2-Chloro-3,5-dimethoxy-4-methylphenyl 28 Cl NH₂ 2-Py2-Bromo-3,5-dimethoxy-4-methylphenyl 29 Cl NH₂ 2-Py2-Iodo-3,5-dimethoxy-4-methylphenyl 30 Cl NH₂ 2-Py2-Fluoro-3,5-dimethoxy-4-methylphenyl 31 Cl NH₂ 2-Py3,4,5-Trimethoxyphenyl 32 Cl NH₂ 2-Py 2-Chloro-3,4,5-trimethoxyphenyl 33Cl NH₂ 2-Py 2-Bromo-3,4,5-trimethoxyphenyl 34 Cl NH₂ 2-Py2-Iodo-3,4,5-trimethoxyphenyl 35 Cl NH₂ 2-Py2-Fluoro-3,4,5-trimethoxyphenyl 36 Cl NH₂ 2-Py 3,4,5-Trimethylphenyl 37Cl NH₂ 2-Py 2-Chloro-3,4,5-trimethylphenyl 38 Cl NH₂ 2-Py2-Bromo-3,4,5-trimethylphenyl 39 Cl NH₂ 2-Py2-Iodo-3,4,5-trimethylphenyl 40 Cl NH₂ 2-Py2-Fluoro-3,4,5-trimethylphenyl 41 Cl NH₂ Et 3,5-Dimethoxy-4-methylphenyl42 Cl NH₂ Et 2-Chloro-3,5-dimethoxy-4-methylphenyl 43 Cl NH₂ Et2-Bromo-3,5-dimethoxy-4-methylphenyl 44 Cl NH₂ Et2-Iodo-3,5-dimethoxy-4-methylphenyl 45 Cl NH₂ Et2-Fluoro-3,5-dimethoxy-4-methylphenyl 46 Cl NH₂ Me3,4,5-Trimethoxyphenyl 47 Cl NH₂ Me 2-Chloro-3,4,5-trimethoxyphenyl 48Cl NH₂ Me 2-Bromo-3,4,5-trimethoxyphenyl 49 Cl NH₂ Me2-Iodo-3,4,5-trimethoxyphenyl 50 Cl NH₂ Me2-Fluoro-3,4,5-trimethoxyphenyl 51 Cl NH₂ Me 3,4,5-Trimethylphenyl 52 ClNH₂ Me 2-Chloro-3,4,5-trimethylphenyl 53 Cl NH₂ Me2-Bromo-3,4,5-trimethylphenyl 54 Cl NH₂ Me 2-Iodo-3,4,5-trimethylphenyl55 Cl NH₂ Me 2-Fluoro-3,4,5-trimethylphenyl 56 Cl NH₂ Me3,5-Dimethoxy-4-methylphenyl 57 Cl NH₂ Me2-Chloro-3,5-dimethoxy-4-methylphenyl 58 Cl NH₂ Me2-Bromo-3,5-dimethoxy-4-methylphenyl 59 Cl NH₂ Me2-Iodo-3,5-dimethoxy-4-methylphenyl 60 Cl NH₂ Me2-Fluoro-3,5-dimethoxy-4-methylphenyl 61 Cl NH₂ Ph3,4,5-Trimethoxyphenyl 62 Cl NH₂ Ph 2-Chloro-3,4,5-trimethoxyphenyl 63Cl NH₂ Ph 2-Bromo-3,4,5-trimethoxyphenyl 64 Cl NH₂ Ph2-Iodo-3,4,5-trimethoxyphenyl 65 Cl NH₂ Ph2-Fluoro-3,4,5-trimethoxyphenyl 66 Cl NH₂ Ph 3,4,5-Trimethylphenyl 67 ClNH₂ Ph 2-Chloro-3,4,5-trimethylphenyl 68 Cl NH₂ Ph2-Bromo-3,4,5-trimethylphenyl 69 Cl NH₂ Ph 2-Iodo-3,4,5-trimethylphenyl70 Cl NH₂ Ph 2-Fluoro-3,4,5-trimethylphenyl 71 Cl NH₂ Ph3,5-Dimethoxy-4-methylphenyl 72 Cl NH₂ Ph2-Chloro-3,5-dimethoxy-4-methylphenyl 73 Cl NH₂ Ph2-Bromo-3,5-dimethoxy-4-methylphenyl 74 Cl NH₂ Ph2-Iodo-3,5-dimethoxy-4-methylphenyl 75 Cl NH₂ Ph2-Fluoro-3,5-dimethoxy-4-methylphenyl 76 Cl NH₂ (CH₂)₂OH3,4,5-Trimethoxyphenyl 77 Cl NH₂ (CH₂)₂OH2-Chloro-3,4,5-trimethoxyphenyl 78 Cl NH₂ (CH₂)₂OH2-Bromo-3,4,5-trimethoxyphenyl 79 Cl NH₂ (CH₂)₂OH2-Iodo-3,4,5-trimethoxyphenyl 80 Cl NH₂ (CH₂)₂OH2-Fluoro-3,4,5-trimethoxyphenyl 81 Cl NH₂ (CH₂)₂OH 3,4,5-Trimethylphenyl82 Cl NH₂ (CH₂)₂OH 2-Chloro-3,4,5-trimethylphenyl 83 Cl NH₂ (CH₂)₂OH2-Bromo-3,4,5-trimethylphenyl 84 Cl NH₂ (CH₂)₂OH2-Iodo-3,4,5-trimethylphenyl 85 Cl NH₂ (CH₂)₂OH2-Fluoro-3,4,5-trimethylphenyl 86 Cl NH₂ (CH₂)₂OH3,5-Dimethoxy-4-methylphenyl 87 Cl NH₂ (CH₂)₂OH2-Chloro-3,5-dimethoxy-4-methylphenyl 88 Cl NH₂ (CH₂)₂OH2-Bromo-3,5-dimethoxy-4-methylphenyl 89 Cl NH₂ (CH₂)₂OH2-Iodo-3,5-dimethoxy-4-methylphenyl 90 Cl NH₂ (CH₂)₂OH2-Fluoro-3,5-dimethoxy-4-methylphenyl 91 Cl NH₂ 4-Py3,4,5-Trimethoxyphenyl 92 Cl NH₂ 4-Py 2-Chloro-3,4,5-trimethoxyphenyl 93Cl NH₂ 4-Py 2-Bromo-3,4,5-trimethoxyphenyl 94 Cl NH₂ 4-Py2-Iodo-3,4,5-trimethoxyphenyl 95 Cl NH₂ 4-Py2-Fluoro-3,4,5-trimethoxyphenyl 96 Cl NH₂ Ph 3,4,5-Trimethylphenyl 97 ClNH₂ Ph 2-Chloro-3,4,5-trimethylphenyl 98 Cl NH₂ Ph2-Bromo-3,4,5-trimethylphenyl 99 Cl NH₂ Ph 2-Iodo-3,4,5-trimethylphenyl100 Cl NH₂ Ph 2-Fluoro-3,4,5-trimethylphenyl 101 Cl NH₂ Ph3,5-Dimethoxy-4-methylphenyl 102 Cl NH₂ Ph2-Chloro-3,5-dimethoxy-4-methylphenyl 103 Cl NH₂ Ph2-Bromo-3,5-dimethoxy-4-methylphenyl 104 Cl NH₂ Ph2-Iodo-3,5-dimethoxy-4-methylphenyl 105 Cl NH₂ Pr2-Fluoro-3,5-dimethoxy-4-methylphenyl 106 Cl NH₂ Pr3,5-Dimethoxy-4-methylphenyl 107 Cl NH₂ Pr2-Chloro-3,5-dimethoxy-4-methylphenyl 108 Cl NH₂ Pr2-Bromo-3,5-dimethoxy-4-methylphenyl 109 Cl NH₂ Pr2-Iodo-3,5-dimethoxy-4-methylphenyl 110 Cl NH₂ Pr2-Fluoro-3,5-dimethoxy-4-methylphenyl 111 Cl NH₂ Pr3,4,5-Trimethoxyphenyl 112 Cl NH₂ Pr 2-Chloro-3,4,5-trimethoxyphenyl 113Cl NH₂ Pr 2-Bromo-3,4,5-trimethoxyphenyl 114 Cl NH₂ Pr2-Iodo-3,4,5-trimethoxyphenyl 115 Cl NH₂ Pr2-Fluoro-3,4,5-trimethoxyphenyl 116 Cl NH₂ Pr 3,4,5-Trimethylphenyl 117Cl NH₂ Pr 2-Chloro-3,4,5-trimethylphenyl 118 Cl NH₂ Pr2-Bromo-3,4,5-trimethylphenyl 119 Cl NH₂ Pr 2-Iodo-3,4,5-trimethylphenyl120 Cl NH₂ Pr 2-Fluoro-3,4,5-trimethylphenyl 121 Cl NH₂ Pr3,5-Dimethoxy-4-methylphenyl 122 Cl NH₂ Pr2-Chloro-3,5-dimethoxy-4-methylphenyl 123 Cl NH₂ Pr2-Bromo-3,5-dimethoxy-4-methylphenyl 124 Cl NH₂ Pr2-Iodo-3,5-dimethoxy-4-methylphenyl 125 Cl NH₂ Pr2-Fluoro-3,5-dimethoxy-4-methylphenyl 126 Cl NH₂ (CH₂)₃OH3,4,5-Trimethoxyphenyl 127 Cl NH₂ (CH₂)₃OH2-Chloro-3,4,5-trimethoxyphenyl 128 Cl NH₂ (CH₂)₃OH2-Bromo-3,4,5-trimethoxyphenyl 129 Cl NH₂ (CH₂)₃OH2-Iodo-3,4,5-trimethoxyphenyl 130 Cl NH₂ (CH₂)₃OH2-Fluoro-3,4,5-trimethoxyphenyl 131 Cl NH₂ (CH₂)₃OH3,4,5-Trimethylphenyl 132 Cl NH₂ (CH₂)₃OH 2-Chloro-3,4,5-trimethylphenyl133 Cl NH₂ (CH₂)₃OH 2-Bromo-3,4,5-trimethylphenyl 134 Cl NH₂ (CH₂)₃OH2-Iodo-3,4,5-trimethylphenyl 135 Cl NH₂ (CH₂)₃OH2-Fluoro-3,4,5-trimethylphenyl 136 Cl NH₂ (CH₂)₃OH3,5-Dimethoxy-4-methylphenyl 137 Cl NH₂ (CH₂)₃OH2-Chloro-3,5-dimethoxy-4-methylphenyl 138 Cl NH₂ (CH₂)₃OH2-Bromo-3,5-dimethoxy-4-methylphenyl 139 Cl NH₂ (CH₂)₃OH2-Iodo-3,5-dimethoxy-4-methylphenyl 140 Cl NH₂ (CH₂)₃OH2-Fluoro-3,5-dimethoxy-4-methylphenyl 141 Cl NH₂ i-Bu3,4,5-Trimethoxyphenyl 142 Cl NH₂ i-Bu 2-Chloro-3,4,5-trimethoxyphenyl143 Cl NH₂ i-Bu 2-Bromo-3,4,5-trimethoxyphenyl 144 Cl NH₂ i-Bu2-Iodo-3,4,5-trimethoxyphenyl 145 Cl NH₂ i-Bu2-Fluoro-3,4,5-trimethoxyphenyl 146 Cl NH₂ i-Bu 3,4,5-Trimethylphenyl147 Cl NH₂ i-Bu 2-Chloro-3,4,5-trimethylphenyl 148 Cl NH₂ i-Bu2-Bromo-3,4,5-trimethylphenyl 149 Cl NH₂ i-Bu2-Iodo-3,4,5-trimethylphenyl 150 Cl NH₂ i-Bu2-Fluoro-3,4,5-trimethylphenyl 151 Cl NH₂ i-Bu3,5-Dimethoxy-4-methylphenyl 152 Cl NH₂ i-Bu2-Chloro-3,5-dimethoxy-4-methylphenyl 153 Cl NH₂ i-Bu2-Bromo-3,5-dimethoxy-4-methylphenyl 154 Cl NH₂ i-Bu2-Iodo-3,5-dimethoxy-4-methylphenyl 155 Cl NH₂ i-Bu2-Fluoro-3,5-dimethoxy-4-methylphenyl 156 Cl NH₂ CN3,4,5-Trimethoxyphenyl 157 Cl NH₂ CN 2-Chloro-3,4,5-trimethoxyphenyl 158Cl NH₂ CN 2-Bromo-3,4,5-trimethoxyphenyl 159 Cl NH₂ CN2-Iodo-3,4,5-trimethoxyphenyl 160 Cl NH₂ CN 3,4,5-Trimethoxyphenyl 161Cl NH₂ CN 2-Chloro-3,4,5-trimethoxyphenyl 162 Cl NH₂ CN2-Bromo-3,4,5-trimethoxyphenyl 163 Cl NH₂ CN2-Iodo-3,4,5-trimethoxyphenyl 164 Cl NH₂ CN2-Fluoro-3,4,5-trimethoxyphenyl 165 Cl NH₂ CN 3,4,5-Trimethylphenyl 166Cl NH₂ CN 2-Chloro-3,4,5-trimethylphenyl 167 Cl NH₂ CN2-Bromo-3,4,5-trimethylphenyl 168 Cl NH₂ CN 2-Iodo-3,4,5-trimethylphenyl169 Cl NH₂ CN 2-Fluoro-3,4,5-trimethylphenyl 170 Cl NH₂ CN3,5-Dimethoxy-4-methylphenyl 171 Cl NH₂ CN2-Chloro-3,5-dimethoxy-4-methylphenyl 172 Cl NH₂ CN2-Bromo-3,5-dimethoxy-4-methylphenyl 173 Cl NH₂ CN2-Iodo-3,5-dimethoxy-4-methylphenyl 174 Cl NH₂ CN2-Fluoro-3,5-dimethoxy-4-methylphenyl 175 Cl NH₂ CH₂OH3,4,5-Trimethoxyphenyl 176 Cl NH₂ CH₂OH 2-Chloro-3,4,5-trimethoxyphenyl177 Cl NH₂ CH₂OH 2-Bromo-3,4,5-trimethoxyphenyl 178 Cl NH₂ CH₂OH2-Iodo-3,4,5-trimethoxyphenyl 179 Cl NH₂ CH₂OH2-Fluoro-3,4,5-trimethoxyphenyl 180 Cl NH₂ CH₂OH 3,4,5-Trimethylphenyl181 Cl NH₂ CH₂OH 2-Chloro-3,4,5-trimethylphenyl 182 Cl NH₂ CH₂OH2-Bromo-3,4,5-trimethylphenyl 183 Cl NH₂ CH₂OH2-Iodo-3,4,5-trimethylphenyl 184 Cl NH₂ CH₂OH2-Fluoro-3,4,5-trimethylphenyl 185 Cl NH₂ CH₂OH3,5-Dimethoxy-4-methylphenyl 186 Cl NH₂ CH₂OH2-Chloro-3,5-dimethoxy-4-methylphenyl 187 Cl NH₂ CH₂OH2-Bromo-3,5-dimethoxy-4-methylphenyl 188 Cl NH₂ CH₂OH2-Iodo-3,5-dimethoxy-4-methylphenyl 189 Cl NH₂ CH₂OH2-Fluoro-3,5-dimethoxy-4-methylphenyl 190 Cl NH₂ i-Pr3,4,5-Trimethoxyphenyl 191 Cl NH₂ i-Pr 2-Chloro-3,4,5-trimethoxyphenyl192 Cl NH₂ i-Pr 2-Bromo-3,4,5-trimethoxyphenyl 193 Cl NH₂ i-Pr2-Iodo-3,4,5-trimethoxyphenyl 194 Cl NH₂ i-Pr2-Fluoro-3,4,5-trimethoxyphenyl 195 Cl NH₂ i-Pr 3,4,5-Trimethylphenyl196 Cl NH₂ i-Pr 2-Chloro-3,4,5-trimethylphenyl 197 Cl NH₂ i-Pr2-Bromo-3,4,5-trimethylphenyl 198 Cl NH₂ i-Pr2-Iodo-3,4,5-trimethylphenyl 199 Cl NH₂ i-Pr2-Fluoro-3,4,5-trimethylphenyl 200 Cl NH₂ i-Pr3,5-Dimethoxy-4-methylphenyl 201 Cl NH₂ i-Pr2-Chloro-3,5-dimethoxy-4-methylphenyl 202 Cl NH₂ i-Pr2-Bromo-3,5-dimethoxy-4-methylphenyl 203 Cl NH₂ i-Pr2-Iodo-3,5-dimethoxy-4-methylphenyl 204 Cl NH₂ i-Pr2-Fluoro-3,5-dimethoxy-4-methylphenyl 205 Cl NH₂ CO₂Et3,4,5-Trimethoxyphenyl 206 Cl NH₂ CO₂Et 2-Chloro-3,4,5-trimethoxyphenyl207 Cl NH₂ CO₂Et 2-Bromo-3,4,5-trimethoxyphenyl 208 Cl NH₂ CO₂Et2-Iodo-3,4,5-trimethoxyphenyl 209 Cl NH₂ CO₂Et2-Fluoro-3,4,5-trimethoxyphenyl 210 Cl NH₂ CO₂Et 3,4,5-Trimethylphenyl211 Cl NH₂ CO₂Et 2-Chloro-3,4,5-trimethylphenyl 212 Cl NH₂ CO₂Et2-Bromo-3,4,5-trimethylphenyl 213 Cl NH₂ CO₂Et2-Iodo-3,4,5-trimethylphenyl 214 Cl NH₂ CO₂Et2-Fluoro-3,4,5-trimethylphenyl 215 Cl NH₂ CO₂Et3,5-Dimethoxy-4-methylphenyl 216 Cl NH₂ CO₂Et2-Chloro-3,5-dimethoxy-4-methylphenyl 217 Cl NH₂ CO₂Et2-Bromo-3,5-dimethoxy-4-methylphenyl 218 Cl NH₂ CO₂Et2-Iodo-3,5-dimethoxy-4-methylphenyl 219 Cl NH₂ CO₂Et2-Fluoro-3,5-dimethoxy-4-methylphenyl 220 Cl NH₂ CH₂—NMe₂3,4,5-Trimethoxyphenyl 221 Cl NH₂ CH₂—NMe₂2-Chloro-3,4,5-trimethoxyphenyl 222 Cl NH₂ CH₂—NMe₂2-Bromo-3,4,5-trimethoxyphenyl 223 Cl NH₂ CH₂—NMe₂2-Iodo-3,4,5-trimethoxyphenyl 224 Cl NH₂ CH₂—NMe₂2-Fluoro-3,4,5-trimethoxyphenyl 225 Cl NH₂ CH₂—NMe₂3,4,5-Trimethylphenyl 226 Cl NH₂ CH₂—NMe₂ 2-Chloro-3,4,5-trimethylphenyl227 Cl NH₂ CH₂—NMe₂ 2-Bromo-3,4,5-trimethylphenyl 228 Cl NH₂ CH₂—NMe₂2-Iodo-3,4,5-trimethylphenyl 229 Cl NH₂ CH₂—NMe₂2-Fluoro-3,4,5-trimethylphenyl 230 Cl NH₂ CH₂—NMe₂3,5-Dimethoxy-4-methylphenyl 231 Cl NH₂ CH₂—NMe₂2-Chloro-3,5-dimethoxy-4-methylphenyl 232 Cl NH₂ CH₂—NMe₂2-Bromo-3,5-dimethoxy-4-methylphenyl 233 Cl NH₂ CH₂—NMe₂2-Iodo-3,5-dimethoxy-4-methylphenyl 234 Cl NH₂ CH₂—NMe₂2-Fluoro-3,5-dimethoxy-4-methylphenyl 235 Cl NH₂ 3-Py3,4,5-Trimethoxyphenyl 236 Cl NH₂ 3-Py 2-Chloro-3,4,5-trimethoxyphenyl237 Cl NH₂ 3-Py 2-Bromo-3,4,5-trimethoxyphenyl 238 Cl NH₂ 3-Py2-Iodo-3,4,5-trimethoxyphenyl 239 Cl NH₂ 3-Py2-Fluoro-3,4,5-trimethoxyphenyl 240 2 Cl NH₂ (CH₂)₂OH3,5-Dimethyl-4-methoxypyridin-2-yl 241 Cl NH₂ (CH₂)₂OH3,5-Dimethyl-4-methoxy-1-oxypyridin-2-yl 242 Cl NH₂ (CH₂)₂OH6-Bromo-3,5-dimethyl-4-methoxypyridin-2-yl 243 Cl NH₂ (CH₂)₂OH6-Chloro-3,5-dimethyl-4-methoxypyridin-2-yl 244 Cl NH₂ (CH₂)₂OH3,5-Dimethyl-4-bromopyridin-2-yl 245 Cl NH₂ (CH₂)₂OH3,5-Dimethyl-4-bromo-1-oxypyridin-2-yl 246 Cl NH₂ (CH₂)₂OH3,5-Dimethyl-4-chloropyridin-2-yl 247 Cl NH₂ (CH₂)₂OH3,5-Dimethyl-4-chloro-1-oxypyridin-2-yl 248 Cl NH₂ (CH₂)₂OH3,5-Dimethyl-4-iodopyridin-2-yl 249 Cl NH₂ (CH₂)₂OH3,5-Dimethyl-4-iodo-1-oxypyridin-2-yl 250 Cl NH₂ (CH₂)₂OH3,5-Dimethyl-4-thiomethyl-pyridin-2-yl 251 Cl NH₂ (CH₂)₂OH3,5-Dimethyl-4-thiomethyl-1-oxypyridin-2-yl 252 Cl NH₂ (CH₂)₂OH3,4,5-Trimethyl-pyridin-2-yl 253 Cl NH₂ (CH₂)₂OH3,4,5-Trimethyl-1-oxypyridin-2-yl 254 Cl NH₂ (CH₂)₂OH4,5,6-Trimethoxypyridin-2-yl 255 Cl NH₂ (CH₂)₂OH4,5,6-Trimethoxy-1-oxypyridin-2-yl 256 Cl NH₂ (CH₂)₂OH3-Bromo-4,5,6-trimethoxypyridin-2-yl 257 Cl NH₂ (CH₂)₂OH3-Chloro-4,5,6-trimethoxypyridin-2-yl 258 Cl NH₂ (CH₂)₂OH3,4,5-Trimethoxy-pyridin-2-yl 259 Cl NH₂ (CH₂)₂OH3,4,5-Trimethoxy-1-oxypyridin-2-yl 260 Cl NH₂ (CH₂)₂OH3-Bromo-3,4,5-trimethoxy-pyridin-2-yl 261 Cl NH₂ (CH₂)₂OH3-Chloro-3,4,5-trimethoxy-pyridin-2-yl 262 Cl NH₂ (CH₂)₂OH4,5,6-Trimethyl-pyridin-2-yl 263 Cl NH₂ (CH₂)₂OH4,5,6-Trimethyl-1-oxypyridin-2-yl 264 Cl NH₂ (CH₂)₂OH4,6-Dimethyl-5-methoxy-pyridin-2-yl 265 Cl NH₂ (CH₂)₂OH4,6-Dimethyl-5-methoxypyridin-3-yl 266 Cl NH₂ (CH₂)₂OH4,6-Dimethyl-5-methoxy-1-oxypyridin-3-yl 267 Cl NH₂ (CH₂)₂OH4,6-Dimethyl-5-bromopyridin-3-yl 268 Cl NH₂ (CH₂)₂OH4,6-Dimethyl-5-chloropyridin-3-yl 269 Cl NH₂ (CH₂)₂OH5,6-Dimethyl-4-bromopyridin-3-yl 270 Cl NH₂ (CH₂)₂OH5,6-Dimethyl-4-chloropyridin-3-yl 271 Cl NH₂ (CH₂)₂OH2,6-Dimethyl-3-methoxypyridin-4-yl 272 Cl NH₂ (CH₂)₂OH2,6-Dimethyl-pyridin-4-yl 273 Cl NH₂ (CH₂)₂OH2,3,6-Trimethyl-pyridin-4-yl 274 Cl NH₂ (CH₂)₂OH2,3,6-Trimethoxy-pyridin-4-yl 275 Cl NH₂ (CH₂)₂OH2,6-Dimethyl-3-bromopyridin-4-yl 276 Cl NH₂ (CH₂)₂OH2,6-Dimethyl-3-chloropyridin-4-yl 277 Cl NH₂ (CH₂)₂OH2,6-Dimethyl-3-methoxy-1-oxy-pyridin-4-yl 278 Cl NH₂ (CH₂)₂OH2,6-Dimethyl-1-oxy-pyridin-4-yl 279 Cl NH₂ (CH₂)₂OH2,3,6-Trimethyl-1-oxy-pyridin-4-yl 280 Cl NH₂ (CH₂)₂OH2,3,6-Trimethoxy-1-oxy-pyridin-4-yl 281 Cl NH₂ (CH₂)₂OH2,6-Dimethyl-3-bromo1-oxy-pyridin-4-yl 282 Cl NH₂ (CH₂)₂OH2,6-Dimethyl-3-chloro1-oxy-pyridin-4-yl 283 Cl NH₂ (CH₂)₂OH4,6-Dimethyl-5-iodopyridin-3-yl 284 Cl NH₂ (CH₂)₂OH3,5-Dimethyl-4-aminopyridin-2-yl 285 Cl NH₂ i-Pr3,5-Dimethyl-4-methoxypyridin-2-yl 286 Cl NH₂ i-Pr3,5-Dimethyl-4-methoxy-1-oxypyridin-2-yl 287 Cl NH₂ i-Pr6-Bromo-3,5-dimethyl-4-methoxypyridin-2-yl 288 Cl NH₂ i-Pr6-Chloro-3,5-dimethyl-4-methoxypyridin-2-yl 289 Cl NH₂ i-Pr3,5-Dimethyl-4-bromopyridin-2-yl 290 Cl NH₂ i-Pr3,5-Dimethyl-4-bromo-1-oxypyridin-2-yl 291 Cl NH₂ i-Pr3,5-Dimethyl-4-chloropyridin-2-yl 292 Cl NH₂ i-Pr3,5-Dimethyl-4-chloro-1-oxypyridin-2-yl 293 Cl NH₂ i-Pr3,5-Dimethyl-4-iodopyridin-2-yl 294 Cl NH₂ i-Pr3,5-Dimethyl-4-iodo-1-oxypyridin-2-yl 295 Cl NH₂ i-Pr3,5-Dimethyl-4-thiomethyl-pyridin-2-yl 296 Cl NH₂ i-Pr3,5-Dimethyl-4-thiomethyl-1-oxypyridin-2-yl 297 Cl NH₂ i-Pr3,4,5-Trimethyl-pyridin-2-yl 298 Cl NH₂ i-Pr3,4,5-Trimethyl-1-oxypyridin-2-yl 299 Cl NH₂ i-Pr4,5,6-Trimethoxypyridin-2-yl 300 Cl NH₂ i-Pr4,5,6-Trimethoxy-1-oxypyridin-2-yl 301 Cl NH₂ i-Pr3-Bromo-4,5,6-trimethoxypyridin-2-yl 302 Cl NH₂ i-Pr3-Chloro-4,5,6-trimethoxypyridin-2-yl 303 Cl NH₂ i-Pr3,4,5-Trimethoxy-pyridin-2-yl 304 Cl NH₂ i-Pr3,4,5-Trimethoxy-1-oxypyridin-2-yl 305 Cl NH₂ i-Pr3-Bromo-3,4,5-trimethoxy-pyridin-2-yl 306 Cl NH₂ i-Pr3-Chloro-3,4,5-trimethoxy-pyridin-2-yl 307 Cl NH₂ i-Pr4,5,6-Trimethyl-pyridin-2-yl 308 Cl NH₂ i-Pr4,5,6-Trimethyl-1-oxypyridin-2-yl 309 Cl NH₂ i-Pr4,6-Dimethyl-5-methoxy-pyridin-2-yl 310 Cl NH₂ i-Pr4,6-Dimethyl-5-methoxypyridin-3-yl 311 Cl NH₂ i-Pr4,6-Dimethyl-5-methoxy-1-oxypyridin-3-yl 312 Cl NH₂ i-Pr4,6-Dimethyl-5-bromopyridin-3-yl 313 Cl NH₂ i-Pr4,6-Dimethyl-5-chloropyridin-3-yl 314 Cl NH₂ i-Pr5,6-Dimethyl-4-bromopyridin-3-yl 315 Cl NH₂ i-Pr5,6-Dimethyl-4-chloropyridin-3-yl 316 Cl NH₂ i-Pr2,6-Dimethyl-3-methoxypyridin-4-yl 317 Cl NH₂ i-Pr2,6-Dimethyl-pyridin-4-yl 318 Cl NH₂ i-Pr 2,3,6-Trimethyl-pyridin-4-yl319 Cl NH₂ i-Pr 2,3,6-Trimethoxy-pyridin-4-yl 320 Cl NH₂ i-Pr2,6-Dimethyl-3-bromopyridin-4-yl 321 Cl NH₂ i-Pr2,6-Dimethyl-3-chloropyridin-4-yl 322 Cl NH₂ i-Pr2,6-Dimethyl-3-methoxy-1-oxy-pyridin-4-yl 323 Cl NH₂ i-Pr2,6-Dimethyl-1-oxy-pyridin-4-yl 324 Cl NH₂ i-Pr2,3,6-Trimethyl-1-oxy-pyridin-4-yl 325 Cl NH₂ i-Pr2,3,6-Trimethoxy-1-oxy-pyridin-4-yl 326 Cl NH₂ i-Pr2,6-Dimethyl-3-bromo1-oxy-pyridin-4-yl 327 Cl NH₂ i-Pr2,6-Dimethyl-3-chloro1-oxy-pyridin-4-yl 328 Cl NH₂ i-Pr4,6-Dimethyl-5-iodopyridin-3-yl 329 Cl NH₂ i-Pr3,5-Dimethyl-4-aminopyridin-2-yl 330 3 Cl NH₂ (CH₂)₃OH3,5-Dimethyl-4-methoxypyridin-2-yl 331 Cl NH₂ (CH₂)₃OH3,5-Dimethyl-4-methoxy-1-oxypyridin-2-yl 332 Cl NH₂ (CH₂)₃OH3,5-Dimethyl-4-bromopyridin-2-yl 333 Cl NH₂ (CH₂)₃OH3,5-Dimethyl-4-bromo-1-oxypyridin-2-yl 334 Cl NH₂ (CH₂)₃OH3,5-Dimethyl-4-chloropyridin-2-yl 335 Cl NH₂ (CH₂)₃OH3,5-Dimethyl-4-chloro-1-oxypyridin-2-yl 336 Cl NH₂ (CH₂)₃OH3,5-Dimethyl-4-iodopyridin-2-yl 337 Cl NH₂ (CH₂)₃OH3,5-Dimethyl-4-iodo-1-oxypyridin-2-yl 338 Cl NH₂ (CH₂)₃OH3,5-Dimethyl-4-thiomethyl-pyridin-2-yl 339 Cl NH₂ (CH₂)₃OH3,5-Dimethyl-4-thiomethyl-1-oxypyridin-2-yl 340 Cl NH₂ (CH₂)₃OH3,4,5-Trimethyl-pyridin-2-yl 341 Cl NH₂ (CH₂)₃OH3,4,5-Trimethyl-1-oxypyridin-2-yl 342 Cl NH₂ (CH₂)₃OH4,5,6-Trimethoxypyridin-2-yl 343 Cl NH₂ (CH₂)₃OH4,5,6-Trimethoxy-1-oxypyridin-2-yl 344 Cl NH₂ (CH₂)₃OH3,4,5-Trimethoxy-pyridin-2-yl 345 Cl NH₂ (CH₂)₃OH3,4,5-Trimethoxy-1-oxypyridin-2-yl 346 Cl NH₂ (CH₂)₃OH4,5,6-Trimethyl-pyridin-2-yl 347 Cl NH₂ (CH₂)₃OH4,5,6-Trimethyl-1-oxypyridin-2-yl 348 Cl NH₂ (CH₂)₃OH4,6-Dimethyl-5-methoxy-pyridin-2-yl 349 Cl NH₂ (CH₂)₃OH4,6-Dimethyl-5-methoxypyridin-3-yl 350 Cl NH₂ (CH₂)₃OH4,6-Dimethyl-5-methoxy-1-oxypyridin-3-yl 351 Cl NH₂ (CH₂)₃OH4,6-Dimethyl-5-bromopyridin-3-yl 352 Cl NH₂ (CH₂)₃OH4,6-Dimethyl-5-chloropyridin-3-yl 353 Cl NH₂ (CH₂)₃OH5,6-Dimethyl-4-bromopyridin-3-yl 354 Cl NH₂ (CH₂)₃OH5,6-Dimethyl-4-chloropyridin-3-yl 355 Cl NH₂ (CH₂)₃OH2,6-Dimethyl-3-methoxypyridin-4-yl 356 Cl NH₂ (CH₂)₃OH2,6-Dimethyl-pyridin-4-yl 357 Cl NH₂ (CH₂)₃OH2,3,6-Trimethyl-pyridin-4-yl 358 Cl NH₂ (CH₂)₃OH2,3,6-Trimethoxy-pyridin-4-yl 359 Cl NH₂ (CH₂)₃OH2,6-Dimethyl-3-bromopyridin-4-yl 360 Cl NH₂ (CH₂)₃OH2,6-Dimethyl-3-chloropyridin-4-yl 361 Cl NH₂ (CH₂)₃OH4,6-Dimethyl-5-iodopyridin-3-yl 362 Cl NH₂ (CH₂)₃OH3,5-Dimethyl-4-aminopyridin-2-yl 363 1 Cl NH₂ CH₂OH3,5-Dimethyl-4-methoxypyridin-2-yl 364 Cl NH₂ CH₂OH3,5-Dimethyl-4-methoxy-1-oxypyridin-2-yl 365 Cl NH₂ CH₂OH3,5-Dimethyl-4-bromopyridin-2-yl 366 Cl NH₂ CH₂OH3,5-Dimethyl-4-bromo-1-oxypyridin-2-yl 367 Cl NH₂ CH₂OH3,5-Dimethyl-4-chloropyridin-2-yl 368 Cl NH₂ CH₂OH3,5-Dimethyl-4-chloro-1-oxypyridin-2-yl 369 Cl NH₂ CH₂OH3,5-Dimethyl-4-iodopyridin-2-yl 370 Cl NH₂ CH₂OH3,5-Dimethyl-4-iodo-1-oxypyridin-2-yl 371 Cl NH₂ CH₂OH3,5-Dimethyl-4-thiomethyl-pyridin-2-yl 372 Cl NH₂ CH₂OH3,5-Dimethyl-4-thiomethyl-1-oxypyridin-2-yl 373 Cl NH₂ CH₂OH3,4,5-Trimethyl-pyridin-2-yl 374 Cl NH₂ CH₂OH3,4,5-Trimethyl-1-oxypyridin-2-yl 375 Cl NH₂ CH₂OH4,5,6-Trimethoxypyridin-2-yl 376 Cl NH₂ CH₂OH4,5,6-Trimethoxy-1-oxypyridin-2-yl 377 Cl NH₂ CH₂OH3,4,5-Trimethoxy-pyridin-2-yl 378 Cl NH₂ CH₂OH3,4,5-Trimethoxy-1-oxypyridin-2-yl 379 Cl NH₂ CH₂OH4,5,6-Trimethyl-pyridin-2-yl 380 Cl NH₂ CH₂OH4,5,6-Trimethyl-1-oxypyridin-2-yl 381 Cl NH₂ CH₂OH4,6-Dimethyl-5-methoxy-pyridin-2-yl 382 Cl NH₂ CH₂OH4,6-Dimethyl-5-methoxypyridin-3-yl 383 Cl NH₂ CH₂OH4,6-Dimethyl-5-methoxy-1-oxypyridin-3-yl 384 Cl NH₂ CH₂OH4,6-Dimethyl-5-bromopyridin-3-yl 385 Cl NH₂ CH₂OH4,6-Dimethyl-5-chloropyridin-3-yl 386 Cl NH₂ CH₂OH5,6-Dimethyl-4-bromopyridin-3-yl 387 Cl NH₂ CH₂OH5,6-Dimethyl-4-chloropyridin-3-yl 388 Cl NH₂ CH₂OH2,6-Dimethyl-3-methoxypyridin-4-yl 389 Cl NH₂ CH₂OH2,6-Dimethyl-pyridin-4-yl 390 Cl NH₂ CH₂OH 2,3,6-Trimethyl-pyridin-4-yl391 Cl NH₂ CH₂OH 2,3,6-Trimethoxy-pyridin-4-yl 392 Cl NH₂ CH₂OH2,6-Dimethyl-3-bromopyridin-4-yl 393 Cl NH₂ CH₂OH2,6-Dimethyl-3-chloropyridin-4-yl 394 Cl NH₂ CH₂OH4,6-Dimethyl-5-iodopyridin-3-yl 395 Cl NH₂ CH₂OH3,5-Dimethyl-4-aminopyridin-2-yl 396 6 Cl NH₂ 2-Py3,5-Dimethyl-4-methoxypyridin-2-yl 397 Cl NH₂ 2-Py3,5-Dimethyl-4-methoxy-1-oxypyridin-2-yl 398 Cl NH₂ 2-Py3,5-Dimethyl-4-bromopyridin-2-yl 399 Cl NH₂ 2-Py3,5-Dimethyl-4-bromo-1-oxypyridin-2-yl 400 Cl NH₂ 2-Py3,5-Dimethyl-4-chloropyridin-2-yl 401 Cl NH₂ 2-Py3,5-Dimethyl-4-chloro-1-oxypyridin-2-yl 402 Cl NH₂ 2-Py3,5-Dimethyl-4-iodopyridin-2-yl 403 Cl NH₂ 2-Py3,5-Dimethyl-4-iodo-1-oxypyridin-2-yl 404 Cl NH₂ 2-Py3,5-Dimethyl-4-thiomethyl-pyridin-2-yl 405 Cl NH₂ 2-Py3,5-Dimethyl-4-thiomethyl-1-oxypyridin-2-yl 406 Cl NH₂ 2-Py3,4,5-Trimethyl-pyridin-2-yl 407 Cl NH₂ 2-Py3,4,5-Trimethyl-1-oxypyridin-2-yl 408 Cl NH₂ 2-Py4,5,6-Trimethoxypyridin-2-yl 409 Cl NH₂ 2-Py4,5,6-Trimethoxy-1-oxypyridin-2-yl 410 Cl NH₂ 2-Py3,4,5-Trimethoxy-pyridin-2-yl 411 Cl NH₂ 2-Py3,4,5-Trimethoxy-1-oxypyridin-2-yl 412 Cl NH₂ 2-Py4,5,6-Trimethyl-pyridin-2-yl 413 Cl NH₂ 2-Py4,5,6-Trimethyl-1-oxypyridin-2-yl 414 Cl NH₂ 2-Py4,6-Dimethyl-5-methoxy-pyridin-2-yl 415 Cl NH₂ 2-Py4,6-Dimethyl-5-methoxypyridin-3-yl 416 Cl NH₂ 2-Py4,6-Dimethyl-5-methoxy-1-oxypyridin-3-yl 417 Cl NH₂ 2-Py4,6-Dimethyl-5-bromopyridin-3-yl 418 Cl NH₂ 2-Py4,6-Dimethyl-5-chloropyridin-3-yl 419 Cl NH₂ 2-Py5,6-Dimethyl-4-bromopyridin-3-yl 420 Cl NH₂ 2-Py5,6-Dimethyl-4-chloropyridin-3-yl 421 Cl NH₂ 2-Py2,6-Dimethyl-3-methoxypyridin-4-yl 422 Cl NH₂ 2-Py2,6-Dimethyl-pyridin-4-yl 423 Cl NH₂ 2-Py 2,3,6-Trimethyl-pyridin-4-yl424 Cl NH₂ 2-Py 2,3,6-Trimethoxy-pyridin-4-yl 425 Cl NH₂ 2-Py2,6-Dimethyl-3-bromopyridin-4-yl 426 Cl NH₂ 2-Py2,6-Dimethyl-3-chloropyridin-4-yl 427 Cl NH₂ 2-Py4,6-Dimethyl-5-iodopyridin-3-yl 428 Cl NH₂ 2-Py3,5-Dimethyl-4-aminopyridin-2-yl 429 Cl NH₂ Ph3,5-Dimethyl-4-methoxypyridin-2-yl 430 Cl NH₂ Ph3,5-Dimethyl-4-methoxy-1-oxypyridin-2-yl 431 Cl NH₂ Ph3,5-Dimethyl-4-bromopyridin-2-yl 432 Cl NH₂ Ph3,5-Dimethyl-4-bromo-1-oxypyridin-2-yl 433 Cl NH₂ Ph3,5-Dimethyl-4-chloropyridin-2-yl 434 Cl NH₂ Ph3,5-Dimethyl-4-chloro-1-oxypyridin-2-yl 435 Cl NH₂ Ph3,5-Dimethyl-4-iodopyridin-2-yl 436 Cl NH₂ Ph3,5-Dimethyl-4-iodo-1-oxypyridin-2-yl 437 Cl NH₂ Ph3,5-Dimethyl-4-thiomethyl-pyridin-2-yl 438 Cl NH₂ Ph3,5-Dimethyl-4-thiomethyl-1-oxypyridin-2-yl 439 Cl NH₂ Ph3,4,5-Trimethyl-pyridin-2-yl 440 Cl NH₂ Ph3,4,5-Trimethyl-1-oxypyridin-2-yl 441 Cl NH₂ Ph4,5,6-Trimethoxypyridin-2-yl 442 Cl NH₂ Ph4,5,6-Trimethoxy-1-oxypyridin-2-yl 443 Cl NH₂ Ph3,4,5-Trimethoxy-pyridin-2-yl 444 Cl NH₂ Ph3,4,5-Trimethoxy-1-oxypyridin-2-yl 445 Cl NH₂ Ph4,5,6-Trimethyl-pyridin-2-yl 446 Cl NH₂ Ph4,5,6-Trimethyl-1-oxypyridin-2-yl 447 Cl NH₂ Ph4,6-Dimethyl-5-methoxy-pyridin-2-yl 448 Cl NH₂ Ph4,6-Dimethyl-5-methoxypyridin-3-yl 449 Cl NH₂ Ph4,6-Dimethyl-5-methoxy-1-oxypyridin-3-yl 450 Cl NH₂ Ph4,6-Dimethyl-5-bromopyridin-3-yl 451 Cl NH₂ Ph4,6-Dimethyl-5-chloropyridin-3-yl 452 Cl NH₂ Ph5,6-Dimethyl-4-bromopyridin-3-yl 453 Cl NH₂ Ph5,6-Dimethyl-4-chloropyridin-3-yl 454 Cl NH₂ Ph2,6-Dimethyl-3-methoxypyridin-4-yl 455 Cl NH₂ Ph2,6-Dimethyl-pyridin-4-yl 456 Cl NH₂ Ph 2,3,6-Trimethyl-pyridin-4-yl 457Cl NH₂ Ph 2,3,6-Trimethoxy-pyridin-4-yl 458 Cl NH₂ Ph2,6-Dimethyl-3-bromopyridin-4-yl 459 Cl NH₂ Ph2,6-Dimethyl-3-chloropyridin-4-yl 460 Cl NH₂ Ph4,6-Dimethyl-5-iodopyridin-3-yl 461 Cl NH₂ Ph3,5-Dimethyl-4-aminopyridin-2-yl 462 Cl NH₂ 3-Py3,5-Dimethyl-4-methoxypyridin-2-yl 463 Cl NH₂ 3-Py3,5-Dimethyl-4-methoxy-1-oxypyridin-2-yl 464 Cl NH₂ 3-Py3,5-Dimethyl-4-bromopyridin-2-yl 465 Cl NH₂ 3-Py3,5-Dimethyl-4-bromo-1-oxypyridin-2-yl 466 Cl NH₂ 3-Py3,5-Dimethyl-4-chloropyridin-2-yl 467 Cl NH₂ 3-Py3,5-Dimethyl-4-chloro-1-oxypyridin-2-yl 468 Cl NH₂ 3-Py3,5-Dimethyl-4-iodopyridin-2-yl 469 Cl NH₂ 3-Py3,5-Dimethyl-4-iodo-1-oxypyridin-2-yl 470 Cl NH₂ 3-Py3,5-Dimethyl-4-thiomethyl-pyridin-2-yl 471 Cl NH₂ 3-Py3,5-Dimethyl-4-thiomethyl-1-oxypyridin-2-yl 472 Cl NH₂ 3-Py3,4,5-Trimethyl-pyridin-2-yl 473 Cl NH₂ 3-Py3,4,5-Trimethyl-1-oxypyridin-2-yl 474 Cl NH₂ 3-Py4,5,6-Trimethoxypyridin-2-yl 475 Cl NH₂ 3-Py4,5,6-Trimethoxy-1-oxypyridin-2-yl 476 Cl NH₂ 3-Py3,4,5-Trimethoxy-pyridin-2-yl 477 Cl NH₂ 3-Py3,4,5-Trimethoxy-1-oxypyridin-2-yl 478 Cl NH₂ 3-Py4,5,6-Trimethyl-pyridin-2-yl 479 Cl NH₂ 3-Py4,5,6-Trimethyl-1-oxypyridin-2-yl 480 Cl NH₂ 3-Py4,6-Dimethyl-5-methoxy-pyridin-2-yl 481 Cl NH₂ 3-Py4,6-Dimethyl-5-methoxypyridin-3-yl 482 Cl NH₂ 3-Py4,6-Dimethyl-5-methoxy-1-oxypyridin-3-yl 483 Cl NH₂ 3-Py4,6-Dimethyl-5-bromopyridin-3-yl 484 Cl NH₂ 3-Py4,6-Dimethyl-5-chloropyridin-3-yl 485 Cl NH₂ 3-Py5,6-Dimethyl-4-bromopyridin-3-yl 486 Cl NH₂ 3-Py5,6-Dimethyl-4-chloropyridin-3-yl 487 Cl NH₂ 3-Py2,6-Dimethyl-3-methoxypyridin-4-yl 488 Cl NH₂ 3-Py2,6-Dimethyl-pyridin-4-yl 489 Cl NH₂ 3-Py 2,3,6-Trimethyl-pyridin-4-yl490 Cl NH₂ 3-Py 2,3,6-Trimethoxy-pyridin-4-yl 491 Cl NH₂ 3-Py2,6-Dimethyl-3-bromopyridin-4-yl 492 Cl NH₂ 3-Py2,6-Dimethyl-3-chloropyridin-4-yl 493 Cl NH₂ 3-Py4,6-Dimethyl-5-iodopyridin-3-yl 494 Cl NH₂ 3-Py3,5-Dimethyl-4-aminopyridin-2-yl 495 Cl NH₂ CH₂—NMe₂3,5-Dimethyl-4-methoxypyridin-2-yl 496 Cl NH₂ CH₂—NMe₂3,5-Dimethyl-4-methoxy-1-oxypyridin-2-yl 497 Cl NH₂ CH₂—NMe₂3,5-Dimethyl-4-bromopyridin-2-yl 498 Cl NH₂ CH₂—NMe₂3,5-Dimethyl-4-bromo-1-oxypyridin-2-yl 499 Cl NH₂ CH₂—NMe₂3,5-Dimethyl-4-chloropyridin-2-yl 500 Cl NH₂ CH₂—NMe₂3,5-Dimethyl-4-chloro-1-oxypyridin-2-yl 501 Cl NH₂ CH₂—NMe₂3,5-Dimethyl-4-iodopyridin-2-yl 502 Cl NH₂ CH₂—NMe₂3,5-Dimethyl-4-iodo-1-oxypyridin-2-yl 503 Cl NH₂ CH₂—NMe₂3,5-Dimethyl-4-thiomethyl-pyridin-2-yl 504 Cl NH₂ CH₂—NMe₂3,5-Dimethyl-4-thiomethyl-1-oxypyridin-2-yl 505 Cl NH₂ CH₂—NMe₂3,4,5-Trimethyl-pyridin-2-yl 506 Cl NH₂ CH₂—NMe₂3,4,5-Trimethyl-1-oxypyridin-2-yl 507 Cl NH₂ CH₂—NMe₂4,5,6-Trimethoxypyridin-2-yl 508 Cl NH₂ CH₂—NMe₂4,5,6-Trimethoxy-1-oxypyridin-2-yl 509 Cl NH₂ CH₂—NMe₂3,4,5-Trimethoxy-pyridin-2-yl 510 Cl NH₂ CH₂—NMe₂3,4,5-Trimethoxy-1-oxypyridin-2-yl 511 Cl NH₂ CH₂—NMe₂4,5,6-Trimethyl-pyridin-2-yl 512 Cl NH₂ CH₂—NMe₂4,5,6-Trimethyl-1-oxypyridin-2-yl 513 Cl NH₂ CH₂—NMe₂4,6-Dimethyl-5-methoxy-pyridin-2-yl 514 Cl NH₂ CH₂—NMe₂4,6-Dimethyl-5-methoxypyridin-3-yl 515 Cl NH₂ CH₂—NMe₂4,6-Dimethyl-5-methoxy-1-oxypyridin-3-yl 516 Cl NH₂ CH₂—NMe₂4,6-Dimethyl-5-bromopyridin-3-yl 517 Cl NH₂ CH₂—NMe₂4,6-Dimethyl-5-chloropyridin-3-yl 518 Cl NH₂ CH₂—NMe₂5,6-Dimethyl-4-bromopyridin-3-yl 519 Cl NH₂ CH₂—NMe₂5,6-Dimethyl-4-chloropyridin-3-yl 520 Cl NH₂ CH₂—NMe₂2,6-Dimethyl-3-methoxypyridin-4-yl 521 Cl NH₂ CH₂—NMe₂2,6-Dimethyl-pyridin-4-yl 522 Cl NH₂ CH₂—NMe₂2,3,6-Trimethyl-pyridin-4-yl 523 Cl NH₂ CH₂—NMe₂2,3,6-Trimethoxy-pyridin-4-yl 524 Cl NH₂ CH₂—NMe₂2,6-Dimethyl-3-bromopyridin-4-yl 525 Cl NH₂ CH₂—NMe₂2,6-Dimethyl-3-chloropyridin-4-yl 526 Cl NH₂ CH₂—NMe₂4,6-Dimethyl-5-iodopyridin-3-yl 527 Cl NH₂ CH₂—NMe₂3,5-Dimethyl-4-aminopyridin-2-yl 528 Cl NH₂ H3,5-Dimethyl-4-methoxypyridin-2-yl 529 Cl NH₂ H3,5-Dimethyl-4-methoxy-1-oxypyridin-2-yl 530 Cl NH₂ H3,5-Dimethyl-4-bromopyridin-2-yl 531 Cl NH₂ H3,5-Dimethyl-4-bromo-1-oxypyridin-2-yl 532 Cl NH₂ H3,5-Dimethyl-4-chloropyridin-2-yl 533 Cl NH₂ H3,5-Dimethyl-4-chloro-1-oxypyridin-2-yl 534 Cl NH₂ H3,5-Dimethyl-4-iodopyridin-2-yl 535 Cl NH₂ H3,5-Dimethyl-4-iodo-1-oxypyridin-2-yl 536 Cl NH₂ H3,5-Dimethyl-4-thiomethyl-pyridin-2-yl 537 Cl NH₂ H3,5-Dimethyl-4-thiomethyl-1-oxypyridin-2-yl 538 Cl NH₂ H3,4,5-Trimethyl-pyridin-2-yl 539 Cl NH₂ H3,4,5-Trimethyl-1-oxypyridin-2-yl 540 Cl NH₂ H4,5,6-Trimethoxypyridin-2-yl 541 Cl NH₂ H4,5,6-Trimethoxy-1-oxypyridin-2-yl 542 Cl NH₂ H3,4,5-Trimethoxy-pyridin-2-yl 543 Cl NH₂ H3,4,5-Trimethoxy-1-oxypyridin-2-yl 544 Cl NH₂ H4,5,6-Trimethyl-pyridin-2-yl 545 Cl NH₂ H4,5,6-Trimethyl-1-oxypyridin-2-yl 546 Cl NH₂ H4,6-Dimethyl-5-methoxy-pyridin-2-yl 547 Cl NH₂ H4,6-Dimethyl-5-methoxypyridin-3-yl 548 Cl NH₂ H4,6-Dimethyl-5-methoxy-1-oxypyridin-3-yl 549 Cl NH₂ H4,6-Dimethyl-5-bromopyridin-3-yl 550 Cl NH₂ H4,6-Dimethyl-5-chloropyridin-3-yl 551 Cl NH₂ H5,6-Dimethyl-4-bromopyridin-3-yl 552 Cl NH₂ H5,6-Dimethyl-4-chloropyridin-3-yl 553 Cl NH₂ H2,6-Dimethyl-3-methoxypyridin-4-yl 554 Cl NH₂ H2,6-Dimethyl-pyridin-4-yl 555 Cl NH₂ H 2,3,6-Trimethyl-pyridin-4-yl 556Cl NH₂ H 2,3,6-Trimethoxy-pyridin-4-yl 557 Cl NH₂ H2,6-Dimethyl-3-bromopyridin-4-yl 558 Cl NH₂ H2,6-Dimethyl-3-chloropyridin-4-yl 559 Cl NH₂ H3,5-Dimethyl-4-aminopyridin-2-yl 560 Cl NH₂ Me3,5-Dimethyl-4-methoxypyridin-2-yl 561 Cl NH₂ Me3,5-Dimethyl-4-methoxy-1-oxypyridin-2-yl 562 Cl NH₂ Me3,5-Dimethyl-4-bromopyridin-2-yl 563 Cl NH₂ Me3,5-Dimethyl-4-bromo-1-oxypyridin-2-yl 564 Cl NH₂ Me3,5-Dimethyl-4-chloropyridin-2-yl 565 Cl NH₂ Me3,5-Dimethyl-4-chloro-1-oxypyridin-2-yl 566 Cl NH₂ Me3,5-Dimethyl-4-iodopyridin-2-yl 567 Cl NH₂ Me3,5-Dimethyl-4-iodo-1-oxypyridin-2-yl 568 Cl NH₂ Me3,5-Dimethyl-4-thiomethyl-pyridin-2-yl 569 Cl NH₂ Me3,5-Dimethyl-4-thiomethyl-1-oxypyridin-2-yl 570 Cl NH₂ Me3,4,5-Trimethyl-pyridin-2-yl 571 Cl NH₂ Me3,4,5-Trimethyl-1-oxypyridin-2-yl 572 Cl NH₂ Me4,6-Dimethyl-5-methoxypyridin-3-yl 573 Cl NH₂ Me4,6-Dimethyl-5-methoxy-1-oxypyridin-3-yl 574 Cl NH₂ Me3,5-Dimethyl-4-aminopyridin-2-yl 575 Cl NH₂ Et3,5-Dimethyl-4-methoxypyridin-2-yl 576 Cl NH₂ Et3,5-Dimethyl-4-methoxy-1-oxypyridin-2-yl 577 Cl NH₂ Et3,5-Dimethyl-4-bromopyridin-2-yl 578 Cl NH₂ Et3,5-Dimethyl-4-bromo-1-oxypyridin-2-yl 579 Cl NH₂ Et3,5-Dimethyl-4-chloropyridin-2-yl 580 Cl NH₂ Et3,5-Dimethyl-4-chloro-1-oxypyridin-2-yl 581 Cl NH₂ Et3,5-Dimethyl-4-iodopyridin-2-yl 582 Cl NH₂ Et3,5-Dimethyl-4-iodo-1-oxypyridin-2-yl 583 Cl NH₂ Et3,5-Dimethyl-4-thiomethyl-pyridin-2-yl 584 Cl NH₂ Et3,5-Dimethyl-4-thiomethyl-1-oxypyridin-2-yl 585 Cl NH₂ Et3,4,5-Trimethyl-pyridin-2-yl 586 Cl NH₂ Et3,4,5-Trimethyl-1-oxypyridin-2-yl 587 Cl NH₂ Et4,6-Dimethyl-5-methoxypyridin-3-yl 588 Cl NH₂ Et4,6-Dimethyl-5-methoxy-1-oxypyridin-3-yl 589 Cl NH₂ Et3,5-Dimethyl-4-aminopyridin-2-yl 590 Cl NH₂ CO₂Et3,5-Dimethyl-4-methoxypyridin-2-yl 591 Cl NH₂ CO₂Et3,5-Dimethyl-4-methoxy-1-oxypyridin-2-yl 592 Cl NH₂ CO₂Et3,5-Dimethyl-4-bromopyridin-2-yl 593 Cl NH₂ CO₂Et3,5-Dimethyl-4-bromo-1-oxypyridin-2-yl 594 Cl NH₂ CO₂Et3,5-Dimethyl-4-chloropyridin-2-yl 595 Cl NH₂ CO₂Et3,5-Dimethyl-4-chloro-1-oxypyridin-2-yl 596 Cl NH₂ CO₂Et3,5-Dimethyl-4-iodopyridin-2-yl 597 Cl NH₂ CO₂Et3,5-Dimethyl-4-iodo-1-oxypyridin-2-yl 598 Cl NH₂ CO₂Et3,5-Dimethyl-4-thiomethyl-pyridin-2-yl 599 Cl NH₂ CO₂Et3,5-Dimethyl-4-thioniethyl-1-oxypyridin-2-yl 600 Cl NH₂ CO₂Et3,4,5-Trimethyl-pyridin-2-yl 601 Cl NH₂ CO₂Et3,4,5-Trimethyl-1-oxypyridin-2-yl 602 Cl NH₂ CO₂Et4,6-Dimethyl-5-methoxypyridin-3-yl 603 Cl NH₂ CO₂Et4,6-Dimethyl-5-methoxy-1-oxypyridin-3-yl 604 Cl NH₂ CO₂Et3,5-Dimethyl-4-aminopyridin-2-yl 605 Cl NH₂ CN3,5-Dimethyl-4-methoxypyridin-2-yl 606 Cl NH₂ CN3,5-Dimethyl-4-methoxy-1-oxypyridin-2-yl 607 Cl NH₂ CN3,5-Dimethyl-4-bromopyridin-2-yl 608 Cl NH₂ CN3,5-Dimethyl-4-bromo-1-oxypyridin-2-yl 609 Cl NH₂ CN3,5-Dimethyl-4-chloropyridin-2-yl 610 Cl NH₂ CN3,5-Dimethyl-4-chloro-1-oxypyridin-2-yl 611 Cl NH₂ CN3,5-Dimethyl-4-iodopyridin-2-yl 612 Cl NH₂ CN3,5-Dimethyl-4-iodo-1-oxypyridin-2-yl 613 Cl NH₂ CN3,5-Dimethyl-4-thiomethyl-pyridin-2-yl 614 Cl NH₂ CN3,5-Dimethyl-4-thiomethyl-1-oxypyridin-2-yl 615 Cl NH₂ CN3,4,5-Trimethyl-pyridin-2-yl 616 Cl NH₂ CN3,4,5-Trimethyl-1-oxypyridin-2-yl 617 Cl NH₂ CN4,6-Dimethyl-5-methoxypyridin-3-yl 618 Cl NH₂ CN4,6-Dimethyl-5-methoxy-1-oxypyridin-3-yl 619 Cl NH₂ CN3,5-Dimethyl-4-aminopyridin-2-yl 620 Cl NH₂ CMe₂OH3,5-Dimethyl-4-methoxypyridin-2-yl 621 Cl NH₂ CMe₂OH3,5-Dimethyl-4-methoxy-1-oxypyridin-2-yl 622 Cl NH₂ CMe₂OH3,5-Dimethyl-4-bromopyridin-2-yl 623 Cl NH₂ CMe₂OH3,5-Dimethyl-4-bromo-1-oxypyridin-2-yl 624 Cl NH₂ CMe₂OH3,5-Dimethyl-4-chloropyridin-2-yl 625 Cl NH₂ CMe₂OH3,5-Dimethyl-4-chloro-1-oxypyridin-2-yl 626 Cl NH₂ CMe₂OH3,5-Dimethyl-4-iodopyridin-2-yl 627 Cl NH₂ CMe₂OH3,5-Dimethyl-4-iodo-1-oxypyridin-2-yl 628 Cl NH₂ CMe₂OH3,5-Dimethyl-4-thiomethyl-pyridin-2-yl 629 Cl NH₂ CMe₂OH3,5-Dimethyl-4-thiomethyl-1-oxypyridin-2-yl 630 Cl NH₂ CMe₂OH3,4,5-Trimethyl-pyridin-2-yl 631 Cl NH₂ CMe₂OH3,4,5-Trimethyl-1-oxypyridin-2-yl 632 Cl NH₂ CMe₂OH4,6-Dimethyl-5-methoxypyridin-3-yl 633 Cl NH₂ CMe₂OH4,6-Dimethyl-5-methoxy-1-oxypyridin-3-yl 634 Cl NH₂ CMe₂OH3,5-Dimethyl-4-aminopyridin-2-yl 635 Cl NH₂ CH₂CMe₂OH3,5-Dimethyl-4-methoxypyridin-2-yl 636 Cl NH₂ CH₂CMe₂OH3,5-Dimethyl-4-methoxy-1-oxypyridin-2-yl 637 Cl NH₂ CH₂CMe₂OH3,5-Dimethyl-4-bromopyridin-2-yl 638 Cl NH₂ CH₂CMe₂OH3,5-Dimethyl-4-bromo-1-oxypyridin-2-yl 639 Cl NH₂ CH₂CMe₂OH3,5-Dimethyl-4-chloropyridin-2-yl 640 Cl NH₂ CH₂CMe₂OH3,5-Dimethyl-4-chloro-1-oxypyridin-2-yl 641 Cl NH₂ CH₂CMe₂OH3,5-Dimethyl-4-iodopyridin-2-yl 642 Cl NH₂ CH₂CMe₂OH3,5-Dimethyl-4-iodo-1-oxypyridin-2-yl 643 Cl NH₂ CH₂CMe₂OH3,5-Dimethyl-4-thiomethyl-pyridin-2-yl 644 Cl NH₂ CH₂CMe₂OH3,5-Dimethyl-4-thiomethyl-1-oxypyridin-2-yl 645 Cl NH₂ CH₂CMe₂OH3,4,5-Trimethyl-pyridin-2-yl 646 Cl NH₂ CH₂CMe₂OH3,4,5-Trimethyl-1-oxypyridin-2-yl 647 Cl NH₂ CH₂CMe₂OH4,6-Dimethyl-5-methoxypyridin-3-yl 648 Cl NH₂ CH₂CMe₂OH4,6-Dimethyl-5-methoxy-1-oxypyridin-3-yl 649 Cl NH₂ CH₂CH₂CMe₂OH3,5-Dimethyl-4-methoxypyridin-2-yl 650 Cl NH₂ CH₂CH₂CMe₂OH3,5-Dimethyl-4-methoxy-1-oxypyridin-2-yl 651 Cl NH₂ CH₂CH₂CMe₂OH3,5-Dimethyl-4-bromopyridin-2-yl 652 Cl NH₂ CH₂CH₂CMe₂OH3,5-Dimethyl-4-bromo-1-oxypyridin-2-yl 653 Cl NH₂ CH₂CH₂CMe₂OH3,5-Dimethyl-4-chloropyridin-2-yl 654 Cl NH₂ CH₂CH₂CMe₂OH3,5-Dimethyl-4-chloro-1-oxypyridin-2-yl 655 Cl NH₂ CH₂CH₂CMe₂OH3,5-Dimethyl-4-iodopyridin-2-yl 656 Cl NH₂ CH₂CH₂CMe₂OH3,5-Dimethyl-4-iodo-1-oxypyridin-2-yl 657 Cl NH₂ CH₂CH₂CMe₂OH3,5-Dimethyl-4-thiomethyl-pyridin-2-yl 658 Cl NH₂ CH₂CH₂CMe₂OH3,5-Dimethyl-4-thiomethyl-1-oxypyridin-2-yl 659 Cl NH₂ CH₂CH₂CMe₂OH3,4,5-Trimethyl-pyridin-2-yl 660 Cl NH₂ CH₂CH₂CMe₂OH3,4,5-Trimethyl-1-oxypyridin-2-yl 661 Cl NH₂ CH₂CH₂CMe₂OH4,6-Dimethyl-5-methoxypyridin-3-yl 662 Cl NH₂ CH₂CH₂CMe₂OH4,6-Dimethyl-5-methoxy-1-oxypyridin-3-yl 663 Cl NH₂

3,5-Dimethyl-4-methoxypyridin-2-yl 664 Cl NH₂

3,5-Dimethyl-4-methoxy-l-oxypyridin-2-yl 665 Cl NH₂

3,5-Dimethyl-4-bromopyridin-2-yl 666 Cl NH₂

3,5-Dimethyl-4-bromo-1-oxypyridin-2-yl 667 Cl NH₂

3,5-Dimethyl-4-chloropyridin-2-yl 668 Cl NH₂

3,5-Dimethyl-4-chloro-1-oxypyridin-2-yl 669 Cl NH₂

3,5-Dimethyl-4-iodopyridin-2-yl 670 Cl NH₂

3,5-Dimethyl-4-iodo-1-oxypyridin-2-yl 671 Cl NH₂

3,5-Dimethyl-4-thiomethyl-pyridin-2yl 672 Cl NH₂

3,5-Dimethyl-4-thiomethyl-1-oxypyridin-2-yl 673 Cl NH₂

3,4,5-Trimethyl-pyridin-2-yl 674 Cl NH₂

3,4,5-Trimethyl-1-oxypyridin-2-yl 675 Cl NH₂

4,6-Dimethyl-5-methoxypyridin-3-yl 676 Cl NH₂

4,6-Dimethyl-5-methoxy-1-oxypyridin-3-yl 677 Cl NH₂

3,5-Dimethyl-4-aminopyridin-2-yl 678 18 Cl NH₂

3,5-Dimethyl-4-methoxypyridin-2-yl 679 Cl NH₂

3,5-Dimethyl-4-methoxy-1-oxypyridin-2-yl 680 Cl NH₂

3,5-Dimethyl-4-bromopyridin-2-yl 681 Cl NH₂

3,5-Dimethyl-4-bromo-1-oxypyridin-2-yl 682 Cl NH₂

3,5-Dimethyl-4-chloropyridin-2-yl 683 Cl NH₂

3,5-Dimethyl-4-chloro-1-oxypyridin-2-yl 684 Cl NH₂

3,5-Dimethyl-4-iodopyridin-2-yl 685 Cl NH₂

3,5-Dimethyl-4-iodo-1-oxypyridin-2-yl 686 Cl NH₂

3,5-Dimethyl-4-thiomethyl-pyridin-2-yl 687 Cl NH₂

3,5-Dimethyl-4-thiomethyl-1-oxypyridin-2-yl 688 Cl NH₂

3,4,5-Trimethyl-pyridin-2-yl 689 Cl NH₂

3,4,5-Trimethyl-1-oxypyridin-2-yl 690 Cl NH₂

4,6-Dimethyl-5-methoxypyridin-3-yl 691 Cl NH₂

4,6-Dimethyl-5-methoxy-1-oxypyridin-3-yl 692 19 Cl NH₂

3,5-Dimethyl-4-methoxypyridin-2-yl 693 Cl NH₂

Dimethyl-4-methoxy-1-oxypyridin-2-yl 694 Cl NH₂

3,5-Dimethyl-4-bromopyridin-2-yl 695 Cl NH₂

3,5-Dimethyl-4-bromo-1-oxypyridin-2-yl 696 Cl NH₂

3,5-Dimethyl-4-chloropyridin-2-yl 697 Cl NH₂

3,5-Dimethyl-4-chloro-1-oxypyridin-2-yl 698 Cl NH₂

3,5-Dimethyl-4-iodopyridin-2-yl 699 Cl NH₂

3,5-Dimethyl-4-iodo-1-oxypyridin-2-yl 700 Cl NH₂

3,5-Dimethyl-4-thiomethyl-pyridin-2-yl 701 Cl NH₂

3,5-Dimethyl-4-thiomethyl-1-oxypyridin-2-yl 702 Cl NH₂

3,4,5-Trimethyl-pyridin-2-yl 703 Cl NH₂

3,4,5-Trimethyl-1-oxypyridin-2-yl 704 Cl NH₂

4,6-Dimethyl-5-methoxypyridin-3-yl 705 Cl NH₂

4,6-Dimethyl-5-methoxy-1-oxypyridin-3-yl 706 Cl NH₂

3,4,5-Trimethoxyphenyl 707 Cl NH₂

2-Chloro-3,4,5-trimethoxyphenyl 708 Cl NH₂

2-Bromo-3,4,5-trimethoxyphenyl 709 Cl NH₂

3,5-Dimethyl--4-methoxyphenyl 710 Cl NH₂

2-Chloro-3,5-Dimethyl-4-methoxyphenyl 711 Cl NH₂

2-Bromo-3,5-Dimethyl-4-methoxyphenyl 712 Cl NH₂

3,4,5-Trimethoxyphenyl 713 Cl NH₂

2-Chloro-3,4,5-trimethoxyphenyl 714 Cl NH₂

2-Bromo-3,4,5-trimethoxyphenyl 715 Cl NH₂

3,5-Dimethyl-4-methoxyphenyl 716 Cl NH₂

2-Chloro-3,5-Dimethyl-4-methoxyphenyl 717 Cl NH₂

2-Bromo-3,5-Dimethyl-4-methoxyphenyl 718 Cl NH₂ (CH₂)₂C(O)NMe₂3,5-Dimethyl-4-methoxypyridin-2-yl 719 Cl NH₂ (CH₂)₂C(O)NMe₂3,5-Dimethyl-4-methoxy-1-oxypyridin-2-yl 720 Cl NH₂ (CH₂)₂C(O)NMe₂3,5-Dimethyl-4-bromopyridin-2-yl 721 Cl NH₂ (CH₂)₂C(O)NMe₂3,5-Dimethyl-4-bromo-1-oxypyridin-2-yl 722 Cl NH₂ (CH₂)₂C(O)NMe₂3,5-Dimethyl-4-chloropyridin-2-yl 723 Cl NH₂ (CH₂)₂C(O)NMe₂3,5-Dimethyl-4-chloro-1-oxypyridin-2-yl 724 Cl NH₂ (CH₂)₂C(O)NMe₂3,5-Dimethyl-4-iodopyridin-2-yl 725 Cl NH₂ (CH₂)₂C(O)NMe₂3,5-Dimethyl-4-iodo-1-oxypyridin-2-yl 726 Cl NH₂ (CH₂)₂C(O)NMe₂3,5-Dimethyl-4-thiomethyl-pyridin-2-yl 727 Cl NH₂ (CH₂)₂C(O)NMe₂3,5-Dimethyl-4-thiomethyl-1-oxypyridin-2-yl 728 Cl NH₂ (CH₂)₂C(O)NMe₂3,4,5-Trimethyl-pyridin-2-yl 729 Cl NH₂ (CH₂)₂C(O)NMe₂3,4,5-Trimethyl-1-oxypyridin-2-yl 730 Cl NH₂ (CH₂)₂C(O)NMe₂4,6-Dimethyl-5-methoxypyridin-3-yl 731 Cl NH₂ (CH₂)₂C(O)NMe₂4,6-Dimethyl-5-methoxy-1-oxypyridin-3-yl 732 11 Cl NH₂

3,5-Dimethyl-4-methoxypyridin-2-yl 733 Cl NH₂

3,5-Dimethyl-4-methoxy-1-oxypyridin-2-yl 734 Cl NH₂

3,5-Dimethyl-4-bromopyridin-2-yl 735 Cl NH₂

3,5-Dimethyl-4-bromo-1-oxypyridin-2-yl 736 Cl NH₂

3,5-Dimethyl-4-chloropyridin-2-yl 737 Cl NH₂

3,5-Dimethyl-4-chloro-1-oxypyridin-2-yl 738 Cl NH₂

3,5-Dimethyl-4-iodopyridin-2-yl 739 Cl NH₂

3,5-Dimethyl-4-iodo-1-oxypyridin-2-yl 740 Cl NH₂

3,5-Dimethyl-4-thiomethyl-pyridin-2-yl 741 Cl NH₂

3,5-Dimethyl-4-thiomethyl-1-oxypyridin-2-yl 742 Cl NH₂

3,4,5-Trimethyl-pyridin-2-yl 743 Cl NH₂

3,4,5-Trimethyl-1-oxypyridin-2-yl 744 Cl NH₂

4,6-Dimethyl-5-methoxypyridin-3-yl 745 Cl NH₂

4,6-Dimethyl-5-methoxy-1-oxypyridin-3-yl 746 12 Cl NH₂ (CH₂)₂CONH₂3,5-Dimethyl-4-methoxypyridin-2-yl 747 Cl NH₂ (CH₂)₂CONH₂3,5-Dimethyl-4-methoxy-1-oxypyridin-2-yl 748 Cl NH₂ (CH₂)₂CONH₂3,5-Dimethyl-4-bromopyridin-2-yl 749 Cl NH₂ (CH₂)₂CONH₂3,5-Dimethyl-4-bromo-1-oxypyridin-2-yl 750 Cl NH₂ (CH₂)₂CONH₂3,5-Dimethyl-4-chloropyridin-2-yl 751 Cl NH₂ (CH₂)₂CONH₂3,5-Dimethyl-4-chloro-1-oxypyridin-2-yl 752 Cl NH₂ (CH₂)₂CONH₂3,5-Dimethyl-4-iodopyridin-2-yl 753 Cl NH₂ (CH₂)₂CONHBOC3,5-Dimethyl-4-methoxypyridin-2-yl 754 Cl NH₂ (CH₂)₂CONHBOC3,5-Dimethyl-4-methoxy-1-oxypyridin-2-yl 755 Cl NH₂ (CH₂)₂CONHBOC6-Bromo-3,5-dimethyl-4-methoxypyridin-2-yl 756 Cl NH₂ (CH₂)₂CONHBOC6-Chloro-3,5-dimethyl-4-methoxypyridin-2-yl 757 Cl NH₂ (CH₂)₂CONHBOC3,5-Dimethyl-4-bromopyridin-2-yl 758 Cl NH₂ (CH₂)₂CONHBOC3,5-Dimethyl-4-bromo-1-oxypyridin-2-yl 759 Cl NH₂ (CH₂)₂CONHBOC3,5-Dimethyl-4-chloropyridin-2-yl 760 Cl NH₂ (CH₂)₂CONHBOC3,5-Dimethyl-4-chloro-1-oxypyridin-2-yl 761 Cl NH₂ (CH₂)₂CONHBOC3,5-Dimethyl-4-iodopyridin-2-yl 762 Cl NH₂ (CH₂)₂CONHBOC3,5-Dimethyl-4-iodo-1-oxypyridin-2-yl 763 Cl NH₂ (CH₂)₂CONHBOC3,5-Dimethyl-4-thiomethyl-pyridin-2-yl 764 Cl NH₂ (CH₂)₂CONHBOC3,5-Dimethyl-4-thiomethyl-1-oxypyridin-2-yl 765 Cl NH₂ (CH₂)₂CONHBOC3,4,5-Trimethyl-pyridin-2-yl 766 Cl NH₂ (CH₂)₂CONHBOC3,4,5-Trimethyl-1-oxypyridin-2-yl 767 Cl NH₂ (CH₂)₂CONHBOC4,5,6-Trimethoxypyridin-2-yl 768 Cl NH₂ (CH₂)₂CONHBOC4,5,6-Trimethoxy-1-oxypyridin-2-yl 769 Cl NH₂ (CH₂)₂CONHBOC3-Bromo-4,5,6-trimethoxypyridin-2-yl 770 Cl NH₂ (CH₂)₂CONHBOC3-Chloro-4,5,6-trimethoxypyridin-2-yl 771 Cl NH₂ (CH₂)₂CONHBOC3,4,5-Trimethoxy-pyridin-2-yl 772 Cl NH₂ (CH₂)₂CONHBOC3,4,5-Trimethoxy-1-oxypyridin-2-yl 773 Cl NH₂ (CH₂)₂CONHBOC3-Bromo-3,4,5-trimethoxy-pyridin-2-yl 774 Cl NH₂ (CH₂)₂CONHBOC3-Chloro-3,4,5-trimethoxy-pyridin-2-yl 775 Cl NH₂ (CH₂)₂CONHBOC4,5,6-Trimethyl-pyridin-2-yl 776 Cl NH₂ (CH₂)₂CONHBOC4,5,6-Trimethyl-1-oxypyridin-2-yl 777 Cl NH₂ (CH₂)₂CONHBOC4,6-Dimethyl-5-methoxy-pyridin-2-yl 778 Cl NH₂ (CH₂)₂CONHBOC4,6-Dimethyl-5-methoxypyridin-3-yl 779 Cl NH₂ (CH₂)₂CONHBOC4,6-Dimethyl-5-methoxy-1-oxypyridin-3-yl 780 Cl NH₂ (CH₂)₂CONHBOC4,6-Dimethyl-5-bromopyridin-3-yl 781 Cl NH₂ (CH₂)₂CONHBOC4,6-Dimethyl-5-chloropyridin-3-yl 782 Cl NH₂ (CH₂)₂CONHBOC5,6-Dimethyl-4-bromopyridin-3-yl 783 Cl NH₂ (CH₂)₂CONHBOC5,6-Dimethyl-4-chloropyridin-3-yl 784 Cl NH₂ (CH₂)₂CONHBOC2,6-Dimethyl-3-methoxypyridin-4-yl 785 Cl NH₂ (CH₂)₂CONHBOC2,6-Dimethyl-pyridin-4-yl 786 Cl NH₂ (CH₂)₂CONHBOC2,3,6-Trimethyl-pyridin-4-yl 787 Cl NH₂ (CH₂)₂CONHBOC2,3,6-Trimethoxy-pyridin-4-yl 788 Cl NH₂ (CH₂)₂CONHBOC2,6-Dimethyl-3-bromopyridin-4-yl 789 Cl NH₂ (CH₂)₂CONHBOC2,6-Dimethyl-3-chloropyridin-4-yl 790 Cl NH₂ (CH₂)₂CONHBOC2,6-Dimethyl-3-methoxy-1-oxy-pyridin-4-yl 791 Cl NH₂ (CH₂)₂CONHBOC2,6-Dimethyl-1-oxy-pyridin-4-yl 792 Cl NH₂ (CH₂)₂CONHBOC2,3,6-Trimethyl-1-oxy-pyridin-4-yl 793 Cl NH₂ (CH₂)₂CONHBOC2,3,6-Trimethoxy-1-oxy-pyridin-4-yl 794 Cl NH₂ (CH₂)₂CONHBOC2,6-Dimethyl-3-bromo1-oxy-pyridin-4-yl 795 Cl NH₂ (CH₂)₂CONHBOC2,6-Dimethyl-3-chloro1-oxy-pyridin-4-yl 796 Cl NH₂ (CH₂)₂CONHBOC4,6-Dimethyl-5-iodopyridin-3-yl 797 Cl NH₂ (CH₂)₂CONHBOC3,5-Dimethyl-4-aminopyridin-2-yl 798 Cl NH₂

3,5-Dimethyl-4-methoxypyridin-2-yl 799 Cl NH₂

3,5-Dimethyl-4-methoxy-1-oxypyridin-2-yl 800 Cl NH₂

6-Bromo-3,5-dimethyl-4-methoxypyridin-2-yl 801 Cl NH₂

6-Chloro-3,5-dimethyl-4-methoxypyridin-2-yl 802 Cl NH₂

3,5-Dimethyl-4-bromopyridin-2-yl 803 Cl NH₂

3,5-Dimethyl-4-bromo-1-oxypyridin-2-yl 804 Cl NH₂

3,5-Dimethyl-4-chloropyridin-2-yl 805 Cl NH₂

3,5-Dimethyl-4-chloro-1-oxypyridin-2-yl 806 Cl NH₂

3,5-Dimethyl-4-iodopyridin-2-yl 807 Cl NH₂

3,5-Dimethyl-4-iodo-1-oxypyridin-2-yl 808 Cl NH₂

3,5-Dimethyl-4-thiomethyl-pyridin-2-yl 809 Cl NH₂

3,5-Dimethyl-4-thiomethyl-1-oxypyridin-2-yl 810 Cl NH₂

3,4,5-Trimethyl-pyridin-2-yl 811 Cl NH₂

3,4,5-Trimethyl-1-oxypyridin-2-yl 812 Cl NH₂

4,5,6-Trimethoxypyridin-2-yl 813 Cl NH₂

4,5,6-Trimethoxy-1-oxypyridin-2-yl 814 Cl NH₂

3-Bromo-4,5,6-trimethoxypyridin-2-yl 815 Cl NH₂

3-Chloro-4,5,6-trimethoxypyridin-2-yl 816 Cl NH₂

3,4,5-Trimethoxy-pyridin-2-yl 817 Cl NH₂

3,4,5-Trimethoxy-1-oxypyridin-2-yl 818 Cl NH₂

3-Bromo-3,4,5-trimethoxy-pyridin-2-yl 819 Cl NH₂

3-Chloro-3,4,5-trimethoxy-pyridin-2-yl 820 Cl NH₂

4,5,6-Trimethyl-pyridin-2-yl 821 Cl NH₂

4,5,6-Trimethyl-1-oxypyridin-2-yl 822 Cl NH₂

4,6-Dimethyl-5-methoxy-pyridin-2-yl 823 Cl NH₂

4,6-Dimethyl-5-methoxypyridin-3-yl 824 Cl NH₂

4,7,6-Dimethyl-5-methoxy-1-oxypyridin-3-yl 825 Cl NH₂

4,6-Dimethyl-5-bromopyridin-3-yl 826 Cl NH₂

4,6-Dimethyl-5-chloropyridin-3-yl 827 Cl NH₂

5,6-Dimethyl-4-bromopyridin-3-yl 828 Cl NH₂

5,6-Dimethyl-4-chloropyridin-3-yl 829 Cl NH₂

2,6-Dimethyl-3-methoxypyridin-4-yl 830 Cl NH₂

2,6-Dimethyl-pyridin-4-yl 831 Cl NH₂

2,3,6-Trimethyl-pyridin-4-yl 832 Cl NH₂

2,3,6-Trimethoxy-pyridin-4-yl 833 Cl NH₂

2,6-Dimethyl-3-bromopyridin-4-yl 834 Cl NH₂

2,6-Dimethyl-3-chloropyridin-4-yl 835 Cl NH₂

2,6-Dimethyl-3-methoxy-1-oxypyridin-4-yl 836 Cl NH₂

2,6-Dimethyl-1-oxy-pyridin-4-yl 837 Cl NH₂

2,3,6-Trimethyl-1-oxypyridin-4-yl 838 Cl NH₂

2,3,6-Trimethoxy-1-oxypyridin-4-yl 839 Cl NH₂

2,6-Dimethyl-3-bromo1-oxypyridin-4-yl 840 Cl NH₂

2,6-Dimethyl-3-chloro1-oxypyridin-4-yl 841 Cl NH₂

4,6-Dimethyl-5-iodopyridin-3-yl 842 Cl NH₂

3,5-Dimethyl-4-aminopyridin-2-yl 843 Cl NH₂

3,5-Dimethyl-4-methoxypyridin-2-yl 844 Cl NH₂

3,5-Dimethyl-4-methoxy-1-oxypyridin-2-yl 845 Cl NH₂

6-Bromo-3,5-dimethyl-4-methoxypyridin-2-yl 846 Cl NH₂

6-Chloro-3,5-dimethyl-4-methoxypyridin-2-yl 847 Cl NH₂

3,5-Dimethyl-4-bromopyridin-2-yl 848 Cl NH₂

3,5-Dimethyl-4-bromo-1-oxypyridin-2-yl 849 Cl NH₂

3,5-Dimethyl-4-chloropyridin-2-yl 850 Cl NH₂

3,5-Dimethyl-4-chloro-1-oxypyridin-2-yl 851 Cl NH₂

3,5-Dimethyl-4-iodopyridin-2-yl 852 Cl NH₂

3,5-Dimethyl-4-iodo-1-oxypyridin-2-yl 853 Cl NH₂

3,5-Dimethyl-4-thiomethyl-pyridin-2-yl 854 Cl NH₂

3,5-Dimethyl-4-thiomethyl-1-oxypyridin-2-yl 855 Cl NH₂

3,4,5-Trimethyl-pyridin-2-yl 856 Cl NH₂

3,4,5-Trimethyl-1-oxypyridin-2-yl 857 Cl NH₂

4,5,6-Trimethoxypyridin-2-yl 858 Cl NH₂

4,5,6-Trimethoxy-1-oxypyridin-2-yl 859 Cl NH₂

3-Bromo-4,5,6-trimethoxypyridin-2-yl 860 Cl NH₂

3-Chloro-4,5,6-trimethoxypyridin-2-yl 861 Cl NH₂

4,6-Dimethyl-5-methoxypyridin-3-yl 862 Cl NH₂

4,6-Dimethyl-5-methoxy-1-oxypyridin-3-yl 863 Br NH₂ (CH₂)OH3,5-Dimethyl-4-methoxypyridin-2-yl 864 Br NH₂ (CH₂)₂OH3,5-Dimethyl-4-methoxypyridin-2-yl 865 Br NH₂ (CH₂)₃OH3,5-Dimethyl-4-methoxypyridin-2-yl 866 Br NH₂

3,5-Dimethyl-4-methoxypyridin-2-yl 867 Br NH₂

3,5-Dimethyl-4-methoxypyridin-2-yl 868 Br NH₂

3,5-Dimethyl-4-methoxypyridin-2-yl 869 Br NH₂ (CH₂)OH3,5-Dimethyl-4-iodopyridin-2-yl 870 Br NH₂ (CH₂)₂OH3,5-Dimethyl-4-iodopyridin-2-yl 871 Br NH₂ (CH₂)₃OH3,5-Dimethyl-4-iodopyridin-2-yl 872 Br NH₂

3,5-Dimethyl-4-iodopyridin-2-yl 873 Br NH₂

3,5-Dimethyl-4-iodopyridin-2-yl 874 Br NH₂

3,5-Dimethyl-4-iodopyridin-2-yl 875 Cl NH₂

3,5-Dimethyl-4-methoxypyridin-2-yl 876 Cl NH₂

2-Chloro-4,5-dimethoxylphenyl 877 Cl NH₂

2-Nitro-4,5-dimethoxylphenyl 878 Cl NH₂

3,4-Dichlorophenyl 879 Cl NH₂

3,5-Dimethoxylphenyl 880 Cl NH₂

2,5-Dimethoxylphenyl 881 Cl NH₂

3,5-Dimethoxylphenyl 882 Cl NH₂

3-Methoxylphenyl 883 Cl NH₂

4-Methoxylphenyl 884 Cl

(CH₂)₂OH 3,5-Dimethyl-4-iodopyridin-2-yl 885 Cl

(CH₂)₃OH 3,5-Dimethyl-4-iodopyridin-2-yl 886 Cl

3,5-Dimethyl-4-iodopyridin-2-yl 887 Cl

3,5-Dimethyl-4-iodopyridin-2-yl 888 Cl

(CH₂)OH 3,5-Dimethyl-4-methoxypyridin-2-yl 889 Cl

(CH₂)₂OH 3,5-Dimethyl-4-methoxypyridin-2-yl 890 Cl

(CH₂)₃OH 3,5-Dimethyl-4-methoxypyridin-2-yl 891 Cl

3,5-Dimethyl-4-methoxypyridin-2-yl 892 Cl

3,5-Dimethyl-4-methoxypyridin-2-yl 893 Cl

3,5-Dimethyl-4-methoxypyridin-2-yl 894 4 Cl NH₂ CH₂N(i-Bu)₂3,5-Dimethyl-4-methoxypyridin-2-yl 895 5 Cl NH₂ CH₂N(i-Bu)₂3,5-Dimethyl-4-methoxypyridin-2-yl 896 7 Cl NH₂ (CH₂)₄OH3,5-Dimethyl-4-methoxypyridin-2-yl 897 8 Cl NH₂ Si(CH₃)₃3,5-Dimethyl-4-methoxypyridin-2-yl 898 9 Cl NH₂ (CH₂)₂CO₂H3,5-Dimethyl-4-methoxypyridin-2-yl 899 10 Cl NH₂ (CH₂)₂CON(Et)₂3,5-Dimethyl-4-methoxypyridin-2-yl 900 13 Cl NH₂ CH₂NHCOOtBu3,5-Dimethyl-4-methoxypyridin-2-yl 901 14 Cl NH₂ CH₂NH₂3,5-Dimethyl-4-methoxypyridin-2-yl 902 Cl NH₂ CH₂NH₂3,5-Dimethyl-4-methoxy-1-oxypyridin-2-yl 903 Cl NH₂ CH₂NH₂3,5-Dimethyl-4-bromopyridin-2-yl 904 Cl NH₂ CH₂NH₂3,5-Dimethyl-4-bromo-1-oxypyridin-2-yl 905 Cl NH₂ CH₂NH₂3,5-Dimethyl-4-chloropyridin-2-yl 906 Cl NH₂ CH₂NH₂3,5-Dimethyl-4-chloro-1-oxypyridin-2-yl 907 Cl NH₂ CH₂NH₂3,5-Dimethyl-4-iodopyridin-2-yl 908 Cl NH₂ CH₂NH₂3,5-Dimethyl-4-iodo-1-oxypyridin-2-yl 909 Cl NH₂ CH₂NH₂3,5-Dimethyl-4-thiomethyl-pyridin-2-yl 910 Cl NH₂ CH₂NH₂3,5-Dimethyl-4-thiomethyl-1-oxypyridin-2-yl 911 Cl NH₂ CH₂NH₂3,4,5-Trimethyl-pyridin-2-yl 912 Cl NH₂ CH₂NH₂3,4,5-Trimethyl-1-oxypyridin-2-yl 913 Cl NH₂ CH₂NH₂4,5,6-Trimethoxypyridin-2-yl 914 Cl NH₂ CH₂NH₂4,5,6-Trimethoxy-1-oxypyridin-2-yl 915 Cl NH₂ CH₂NH₂3,4,5-Trimethoxy-pyridin-2-yl 916 Cl NH₂ CH₂NH₂3,4,5-Trimethoxy-1-oxypyridin-2-yl 917 Cl NH₂ CH₂NH₂4,5,6-Trimethyl-pyridin-2-yl 918 Cl NH₂ CH₂NH₂4,5,6-Trimethyl-1-oxypyridin-2-yl 919 Cl NH₂ CH₂NH₂4,6-Dimethyl-5-methoxy-pyridin-2-yl 920 Cl NH₂ CH₂NH₂4,6-Dimethyl-5-methoxypyridin-3-yl 921 Cl NH₂ CH₂NH₂4,6-Dimethyl-5-methoxy-1-oxypyridin-3-yl 922 Cl NH₂ CH₂NH₂4,6-Dimethyl-5-bromopyridin-3-yl 923 Cl NH₂ CH₂NH₂4,6-Dimethyl-5-chloropyridin-3-yl 924 Cl NH₂ CH₂NH₂5,6-Dimethyl-4-bromopyridin-3-yl 925 Cl NH₂ CH₂NH₂5,6-Dimethyl-4-chloropyridin-3-yl 926 Cl NH₂ CH₂NH₂2,6-Dimethyl-3-methoxypyridin-4-yl 927 Cl NH₂ CH₂NH₂2,6-Dimethyl-pyridin-4-yl 928 Cl NH₂ CH₂NH₂ 2,3,6-Trimethyl-pyridin-4-yl929 Cl NH₂ CH₂NH₂ 2,3,6-Trimethoxy-pyridin-4-yl 930 Cl NH₂ CH₂NH₂2,6-Dimethyl-3-bromopyridin-4-yl 931 Cl NH₂ CH₂NH₂2,6-Dimethyl-3-chloropyridin-4-yl 932 Cl NH₂ CH₂NH₂4,6-Dimethyl-5-iodopyridin-3-yl 933 Cl NH₂ CH₂NH₂3,5-Dimethyl-4-aminopyridin-2-yl 934 15 Cl NH₂ (CH₂)₃NHCOOtBu3,5-Dimethyl-4-methoxypyridin-2-yl 935 16 Cl NH₂ (CH₂)₃NH₂3,5-Dimethyl-4-methoxypyridin-2-yl 936 17 Cl NH₂

3,5-Dimethyl-4-methoxypyridin-2-yl 937 20 Cl NH₂ (CH₂)₂NHtBu3,5-Dimethyl-4-methoxypyridin-2-yl 938 21 Cl NH₂ (CH₂)₂NHtBu3,5-Dimethyl-4-methoxypyridin-2-yl 939 22 Cl NH₂ CH₂OCO CH₂NMe₂3,5-Dimethyl-4-methoxypyridin-2-yl 940 Cl NH₂ CH₂CH(OH)CH₃3,5-Dimethyl-4-methoxypyridin-2-yl 941 Cl NH₂ CH₂C(O)CH₃3,5-Dimethyl-4-methoxypyridin-2-yl wherein each abbreviation has itsusual meaning, which would be known to one of skill in the chemicalarts, i.e. Me = methyl; Et = ethyl; Pr = propyl; i-Pr = iso-propyl; Bu =butyl; i-Bu = iso-butyl; tBu = tert-butyl; Ph = phenyl; BOC =tert-butoxycarbonyl; 2-Py = 2-pyridyl; 3-Py = 3-pyridyl; 4-Py =4-pyridyl.

Selected compounds in TABLE 1 are compounds 2, 3, 4, 7, 8, 12, 13, 17,18, 22, 23, 27, 28, 32, 33, 38, 42, 43, 47, 48, 52, 53, 57, 58, 62, 63,67, 68, 72, 73, 77, 78, 82, 83, 87, 88, 92, 93, 102, 103, 107, 108, 112,113, 117, 118, 122, 123, 127, 128, 132, 133, 137, 138, 147, 148, 152,153, 157, 158, 161, 162, 166, 167, 171, 172, 176, 177, 181, 182, 186,187, 191, 192, 196, 197, 201, 202, 206, 207, 211, 212, 216, 217, 221,222, 226, 227, 231, 232, 236, 237, 240, 241, 244, 246, 248, 265, 266,285, 289, 291, 293, 310, 311, 330, 331, 332, 334, 349, 350, 363, 364,365, 367, 369, 382, 383, 396, 397, 398, 400, 402, 415, 416, 429, 430,431, 433, 435, 448, 449, 462, 463, 464, 466, 468, 481, 482, 495, 496,497, 499, 501, 514, 515, 528, 529, 530, 532, 534, 547, 548, 560, 561,652, 566, 587, 588, 590, 591, 592, 594, 596, 602, 603, 620, 621, 622,624, 626, 632, 633, 635, 636, 637, 639, 641, 661, 662, 663, 664, 665,667, 669, 675, 676, 678, 679, 680, 682, 684, 690, 691, 692, 693, 694,696, 698, 704, 705, 707, 708, 710, 711, 713, 714, 716, 717, 718, 719,720, 722, 730, 731, 732, 733, 734, 736, 738, 744, 745, 746, 747, 748,750, 752, 753, 754, 757, 759, 761, 778, 779, 782, 783, 784, 798, 799,802, 804, 806, 823, 824, 843, 844, 847, 849, 851, 861, 862, 863, 864,865, 866, 867, 868, 869, 870, 875, 894, 895, 896, 899, 900, 901, 903,905, 907, 920, 921, 934, 935, 936, 937, 938, 939, 940, and 941. Furtherselected compounds are 77, 78, 82, 83, 87, 88, 92, 93, 240, 241, 248,265, 285, 293, 310, 330, 349, 382, 396, 415, 429, 448, 462, 481, 495,514, 528, 560, 587, 602, 620, 632, 635, 661, 663, 675, 678, 690, 692,704, 718, 730, 732, 744, 746, 753, 778, 798, 823, 863, 864, 865, 866,867, 868, 869, 875, 894, 895, 896, 899, 900, 901, 903, 905, 920, 934,935, 936, 940, and 941.

Compounds of the invention exhibit improved HSP90 inhibiting activitiesover some pyrrolo[2,3-d]pyrimidine compounds of Formula II:

wherein R^(3′) is not alkynyl, which were disclosed in U.S. patentapplication Ser. No. 10/945,851 and PCT Application US04/31248. Thepyrrolopyrimidine compounds specified in the EXAMPLE sections of theabove patent applications carry either no substituent or a substitutedalkyl group. Compound 0, one of the embodiments of the compounds ofFormula II, carries no substitutents on the C-5 position (ie R^(3′)═H)and it has an IC₅₀=98 nM as measured by a Her-2 degradation assay(described in EXAMPLE section below). A simple alkyl or a substitutedalkyl at C-5 (ie R³=alkyl or substituted alkyl) can only bring two-foldadditional activity. Conversely, if C-5 is alkyne substituted, thepotency increases approximately 10-20 fold. (See, Series A: compounds A,B, C and D below).

For example, the alkynyl substituents at the C-5 position (see Series B:Cpds E, F, G and H), showed an potency range of 6-17 nM an improvementof at least tenfold over the parent Compound 0. Further, when twocompounds having exactly the same structure, except for the presence ofa triple vs. single bond, e.g. comparing Compound D to Compound E,Compound E which has a propargylic alcohol substituted on C-5 has anIC₅₀=17 nM, while Compound D which has a propanol substituent on C-5 hasan IC₅₀=38 mM, a two-fold improvement. The difference in potency indexbetween alkynes and non-alkynes measured by a secondary assay is alsostriking. The secondary assay measures the efficiency of killing tumorcells. For a given compound, the potency index is defined as:Index=EC50(compound)/EC₅₀(control)where the control is 17-AAG (17-allyl-17-desmethoxy-geldanamycin). EC₅₀is defined as the amount of compound added to effect a 50% reduction inviable cell number. Selected compounds were tested on cells of MCF7 andBT474 breast tumor cell lines. The assay shows that Compound 0 is over10 times less active than 17-AAG. The best non-alkyne substitutedanalogs are 6 to >50 times less potent than 17-AAG. In contrast, thealkyne substituted analogs can be more potent than 17-AAG (see Cpd. H)—a record in the field of HSP90 inhibitors, or nearly as active, with anindex between 0.7-6. TABLE 2 summarized the result of the assay.

TABLE 2 Potency Index against Tumor Cells from MCF7 and BT474 Cell LinesSubsitutent MCF7 Index BT474 Index Compound @ C-5 (μM) (μM) 0 none 0.40.3 A non-alkyne 0.2 1.0 B non-alkyne 0.08 0.3 C non-alkyne 0.6 0.3 Dnon-alkyne 0.23 1.0 E alkyne 0.13 0.02 F alkyne 0.006 0.13 G alkyne 0.030.02 H alkyne 0.01 0.1

Synthesis of the Compounds of the Invention

The compounds of the present invention may be prepared from5-halo-pyrolo[2,3-d]pyrimidines according to Scheme A below. Thepreparation of the starting material,N-(4-chloro-5-iodo-7H-pyrrolo[2,3-d]pyrimidin-2-yl)pivalamide, whereinPG is the pivaloyl protecting group and X═I, has been reported in Seela,F. Synthesis 2004, 8, 1203 and references therein.

Treatment ofN-(4-chloro-5-iodo-7H-pyrrolo[2,3-d]pyrimidin-2-yl)pivalaride with theappropriate alkylating agent in a polar solvent (e.g. DMF, DMSO, NMP),in the presence of a base (e.g. K₂CO₃, C₂CO₃, NaH, NaOH, t-BuOH) affordsthe N(7)-alkylated adduct. The alkylation is typically run at 20-60° C.for 0.5-24 h.

Cleavage of the NH₂ protecting group gives the 2-amino derivative If theprotecting group is a pivaloyl group, it can be cleaved with ZnCl₂. Thedepivaloylation is typically run in EtOH, with 1-10 vol % of water, at5085° C., for 2-24 h. Zinc (IB) chloride can be substituted with otherLewis acids, such as ZnI₂ or CuCl which, however, may not give yields ashigh. Sonogasira coupling of4-chloro-7-alkyl-5-halo-7H-pyrrolo[2,3-d]pyrimidin-2-amine with analkyne of Formula HC≡C—R¹ gives the desired alkyne. Typical reactionconditions require Pd(PPh₃)₄/CuI as catalytic system, Et₃N as base, andDCM or DMF as solvent. The reactions are typically run at 20-50° C. for0.5 to 24 h. However, a variety of alternative catalyticsystems/base/conditions can be used (see Liang, B. et. al., J. Org.Chem. 2005, 70, 391 and references therein).

The R group can be further manipulated if necessary, as illustratedbelow:

For illustration purposes, the propargylic alcohol can be converted tothe mesylate or bromide using methods well known in the art. The mesylor bromide group can then be displaced by nucleophiles such as amines.Care must be taken to avoid the unwanted nucleophilic substitution ofthe 4-Cl atom.

The sequence of these steps can be performed in a different order thanthe one indicated on the Scheme, as for instance (i) alkylation, (ii),Sonogashira coupling, and (iii) deprotection.

Pharmaceutical Compositions, Medicaments, Dosaging and Modes ofAdministration The present invention is also directed to the use ofalkynyl pyrrolo[2,3-d]pyrimidine compounds of Formula I and theirrelated analogs, and their polymorphs, solvates, esters, tautomers,diastereomers, enantiomers, pharmaceutically acceptable salts andprodrugs thereof. In some embodiments, the compounds are used for thetreatment or prevention of diseases that are HSP90-dependent. In someembodiments, the compounds are used in the manufacture of a medicament.In other embodiments, the compounds are used in the manufacture of amedicament for the therapeutic and/or prophylactic treatment of diseasesand conditions that are HSP90-dependent. Examples of such diseases andconditions include disorders such as inflammatory diseases, infections,autoimmune disorders, stroke, ischemia, cardiac disorder, neurologicaldisorders, fibrogenic disorders, proliferative disorders, tumors,leukemias, chronic lymphocytic leukemia, acquired immunodeficiencysyndrome, neoplasms, cancers, carcinomas, metabolic diseases, andmalignant disease. The fibrogenic disorders include but are not limitedto scleroderma, polymyositis, systemic lupus, rheumatoid arthritis,liver cirrhosis, keloid formation, interstitial nephritis and pulmonaryfibrosis.

The present invention features pharmaceutical compositions andmedicaments comprising the compound of Formula I, or a polymorph,solvate, ester, tautomer, enantiomer, diastereomer, pharmaceuticallyacceptable salt thereof, or prodrug thereof, of any of the precedingaspects and embodiments and one or more pharmaceutical excipients. Thoseof ordinary skill in the art are familiar with formulation andadministration techniques that can be employed with the compounds andmethods of the invention, e.g., as discussed in Goodman and Gilman, ThePharmacological Basis of Therapeutics (10th edition); Pergamon; andRemington's, Pharmaceutical Sciences (20th edition), Mack PublishingCo., Easton, Pa.

The compounds utilized in the methods of the instant invention may beadministered either alone or in combination with pharmaceuticallyacceptable carriers, excipients or diluents, in a pharmaceuticalcomposition, according to standard pharmaceutical practice. Thecompounds can be administered orally or parenterally, including theintravenous, intramuscular, intraperitoneal, subcutaneous, rectal andtopical routes of administration. For example, the therapeutic orpharmaceutical compositions of the invention can be administered locallyto the area in need of treatment. This may be achieved by, for example,but not limited to, local infusion during surgery, topical application,e.g., cream, ointment, injection, catheter, or implant, said implantmade, e.g., out of a porous, non-porous, or gelatinous material,including membranes, such as sialastic membranes, or fibers. Theadministration can also be by direct injection at the site (or formersite) of a tumor or neoplastic or pre-neoplastic tissue. Still further,the compounds or compositions of the invention can be delivered in avesicle, e.g., a liposome (see, for example, Langer, Science 1990, 249,1527-1533; Treat et al., Liposomes in the Therapy of Infectious Diseaseand Cancer, Lopez-Bernstein and Fidler, Ed., Liss, N.Y., pp. 353-365,1989).

The compounds and pharmaceutical compositions used in the methods of thepresent invention can also be delivered in a controlled release system.In one embodiment, a pump may be used (see, Sefton, 1987, CRC Crit. Ref.Biomed. Eng. 14:201; Buchwald et al. Surgery, 1980 88, 507; Saudek etal. N. Engl. J. Med. 1989, 321, (574). Additionally, a controlledrelease system can be placed in proximity of the therapeutic target.(See, Goodson, Medical Applications of Controlled Release, 1984, Vol. 2,pp. 115-138).

The pharmaceutical compositions used in the methods of the instantinvention can also contain the active ingredient in a form suitable fororal use, for example, as tablets, troches, lozenges, aqueous or oilysuspensions, dispersible powders or granules, emulsions, hard or softcapsules, or syrups or elixirs. Compositions intended for oral use maybe prepared according to any method known in the art for the manufactureof pharmaceutical compositions, and such compositions may contain one ormore agents selected from the group consisting of sweetening agents,flavoring agents, coloring agents and preserving agents in order toprovide pharmaceutically elegant and palatable preparations. Tabletscontain the active ingredient in admixture with non-toxicpharmaceutically acceptable excipients which are suitable for themanufacture of tablets. These excipients may be, for example, inertdiluents, such as calcium carbonate, sodium carbonate, lactose, calciumphosphate or sodium phosphate; granulating and disintegrating agents,such as miicrocrystalline cellulose, sodium crosscarmellose, cornstarch, or alginic acid; binding agents, for example starch, gelatin,polyvinyl-pyrrolidone or acacia, and lubricating agents, for example,magnesium stearate, stearic acid or talc. The tablets may be un-coatedor coated by known techniques to mask the taste of the drug or delaydisintegration and absorption in the gastrointestinal tract and therebyprovide a sustained action over a longer period. For example, a watersoluble taste masking material such as hydroxypropylmethyl-cellulose orhydroxypropylcellulose, or a time delay material such as ethylcellulose, or cellulose acetate butyrate may be employed as appropriate.

Formulations for oral use may also be presented as hard gelatin capsuleswherein the active ingredient is mixed with an inert solid diluent, forexample, calcium carbonate, calcium phosphate or kaolin, or as softgelatin capsules wherein the active ingredient is mixed with watersoluble carrier such as polyethyleneglycol or an oil medium, for examplepeanut oil, liquid paraffin, or olive oil.

Aqueous suspensions contain the active material in admixture withexcipients suitable for the manufacture of aqueous suspensions. Suchexcipients are suspending agents, for example sodiumcarboxymethylcellulose, methylcellulose, hydroxypropylmethyl-cellulose,sodium alginate, polyvinyl-pyrrolidone, gum tragacanth and gum acacia;dispersing or wetting agents may be a naturally-occurring phosphatide,for example lecithin, or condensation products of an alkylene oxide withfatty acids, for example polyoxyethylene stearate, or condensationproducts of ethylene oxide with long chain aliphatic alcohols, forexample heptadecaethylene-oxycetanol, or condensation products ofethylene oxide with partial esters derived from fatty acids and ahexitol such as polyoxyethylene sorbitol monooleate, or condensationproducts of ethylene oxide with partial esters derived from fatty acidsand hexitol anhydrides, for example polyethylene sorbitan monooleate.The aqueous suspensions may also contain one or more preservatives, forexample ethyl, or n-propyl p-hydroxybenzoate, one or more coloringagents, one or more flavoring agents, and one or more sweetening agents,such as sucrose, saccharin or aspartame.

Oily suspensions may be formulated by suspending the active ingredientin a vegetable oil, for example arachis oil, olive oil, sesame oil orcoconut oil, or in mineral oil such as liquid paraffin. The oilysuspensions may contain a thickening agent, for example beeswax, hardparaffin or cetyl alcohol. Sweetening agents such as those set forthabove, and flavoring agents may be added to provide a palatable oralpreparation. These compositions may be preserved by the addition of ananti-oxidant such as butylated hydroxyanisol or alpha-tocopherol.

Dispersible powders and granules suitable for preparation of an aqueoussuspension by the addition of water provide the active ingredient inadmixture with a dispersing or wetting agent, suspending agent and oneor more preservatives. Suitable dispersing or wetting agents andsuspending agents are exemplified by those already mentioned above.Additional excipients, for example sweetening, flavoring and coloringagents, may also be present. These compositions may be preserved by theaddition of an anti-oxidant such as ascorbic acid.

The compounds and pharmaceutical compositions used in the methods of theinstant invention may also be in the form of an oil-in-water emulsion.The oily phase may be a vegetable oil, for example olive oil or arachisoil, or a mineral oil, for example liquid paraffin, or mixtures ofthese. Suitable emulsifying agents may be naturally-occurringphosphatides, for example soybean lecithin, and esters or partial estersderived from fatty acids and hexitol anhydrides, for example sorbitanmonooleate, and condensation products of the said partial esters withethylene oxide, for example polyoxyethylene sorbitan monooleate. Theemulsions may also contain sweetening agents, flavoring agents,preservatives and antioxidants.

Syrups and elixirs may be formulated with sweetening agents, for exampleglycerol, propylene glycol, sorbitol or sucrose. Such formulations mayalso contain a demulcent, a preservative, flavoring and coloring agentsand antioxidant.

The pharmaceutical compositions may be in the form of a sterileinjectable aqueous solution. Among the acceptable vehicles and solventsthat may be employed are water, Ringer's solution and isotonic sodiumchloride solution.

The sterile injectable preparation may also be a sterile injectableoil-in-water microemulsion where the active ingredient is dissolved inthe oily phase. For example, the active ingredient may be firstdissolved in a mixture of soybean oil and lecithin. The oil solution maythen be introduced into a water and glycerol mixture and processed toform a microemulsion.

The injectable solutions or microemulsions may be introduced into apatient's blood-stream by local bolus injection. Alternatively, it maybe advantageous to administer the solution or microemulsion in such away as to maintain a constant circulating concentration of the instantcompound. In order to maintain such a constant concentration, acontinuous intravenous delivery device may be utilized. An example ofsuch a device is the Deltec CADD-PLUS™ model 5400 intravenous pump.

The pharmaceutical compositions may be in the form of a sterileinjectable aqueous or oleagenous suspension for intramuscular andsubcutaneous administration. This suspension may be formulated accordingto the known art using those suitable dispersing or wetting agents andsuspending agents which have been mentioned above, The sterileinjectable preparation may also be a sterile injectable solution orsuspension in a non-toxic parenterally-acceptable diluent or solvent,for example as a solution in 1,3-butanediol. In addition, sterile, fixedoils are conventionally employed as a solvent or suspending medium. Forthis purpose any bland fixed oil may be employed including syntheticmono- or diglycerides. In addition, fatty acids such as oleic acid finduse in the preparation of injectables.

The compounds of the present invention used in the methods of thepresent invention may also be administered in the form of suppositoriesfor rectal administration of the drug. These compositions can beprepared by mixing the inhibitors with a suitable non-irritatingexcipient which is solid at ordinary temperatures but liquid at therectal temperature and will therefore melt in the rectum to release thedrug. Such materials include cocoa butter, glycerinated gelatin,hydrogenated vegetable oils, mixtures of polyethylene glycols of variousmolecular weights and fatty acid esters of polyethylene glycol. Fortopical use, creams, ointments, jellies, solutions or suspensions, etc.,containing a compound or composition of the invention can be used. Asused herein, topical application can include mouth washes and gargles.

The compounds used in the methods of the present invention can beadministered in intranasal form via topical use of suitable intranasalvehicles and delivery devices, or via transdermal routes, using thoseforms of transdermal skin patches well known to those of ordinary skillin the art. To be administered in the form of a transdermal deliverysystem, the dosage administration will, of course, be continuous ratherthan intermittent throughout the dosage regimen.

The methods, compounds and compositions of the instant invention mayalso be used in conjunction with other well known therapeutic agentsthat are selected for their particular usefulness against the conditionthat is being treated. For example, the instant compounds may be usefulin combination with known anti-cancer and cytotoxic agents. Further, theinstant methods and compounds may also be useful in combination withother inhibitors of parts of the signaling pathway that links cellsurface growth factor receptors to nuclear signals initiating cellularproliferation.

The methods of the present invention may also be useful with otheragents that inhibit angiogenesis and thereby inhibit the growth andinvasiveness of tumor cells, including, but not limited to VEGF receptorinhibitors, including ribozymes and antisense targeted to VEGFreceptors, angiostatin and endostatin. Examples of antineoplastic agentsthat can be used in combination with the compounds and methods of thepresent invention include, in general, and as appropriate, alkylatingagents, anti-metabolites, epidophyllotoxins, an antineoplastic enzyme, atopoisomerase inhibitor, procarbazine, mitoxantrone, platinumcoordination complexes, biological response modifiers and growthinhibitors, hormonal/anti-hormonal therapeutic agents and haematopoieticgrowth factors. Exemplary classes of antineoplastic include theanthracyclines, vinca drugs, mitomycins, bleomycins, cytotoxicnucleosides, epothilones, discodermolide, pteridines, diynenes andpodophyllotoxins. Particularly useful members of those classes include,for example, caminomycin, daunorubicin, aminopterin, methotrexate,methopterin, dichloromethotrexate, mitomycin C, porfiromycin,5-fluorouracil, 6-mercaptopurine, gemcitabine, cytosine arabinoside,podophyllotoxin or podo-phyllotoxin derivatives such as etoposide,etoposide phosphate or teniposide, melphalan, vinblastine, vincristine,leurosidine, vindesine, leurosine, paclitaxel and the like. Other usefulantineoplastic agents include estramustine, carboplatin,cyclophosphamide, bleomycin, gemcitibine, ifosamide, melphalan,hexamethyl melamine, thiotepa, cytarabin, idatrexate, trimetrexate,dacarbazine, L-asparaginase, camptothecin, CPT-11, topotecan, ara-C,bicalutamide, flutamide, leuprolide, pyridobenzoindole derivatives,interferons and interleukins.

When a compound or composition of the invention is administered into ahuman subject, the daily dosage will normally be determined by theprescribing physician with the dosage generally varying according to theage, weight, and response of the individual patient, as well as theseverity of the patient's symptoms. In one exemplary application, asuitable amount of compound is administered to a mammal undergoingtreatment for cancer, for example, breast cancer. Administrationtypically occurs in an amount of between about 0.01 mg/kg of body weightto about 100 mg/kg of body weight per day (administered in single ordivided doses), more preferably at least about 0.1 mg/kg of body weightper day. A particular therapeutic dosage can include, e.g., from about0.01 mg to about 1000 mg of compound, and preferably includes, e.g.,from about 1 mg to about 1000 mg. The quantity of active compound in aunit dose of preparation may be varied or adjusted from about 0.1 mg to1000 mg, preferably from about 1 mg to 300 mg, more preferably 10 mg to200 mg, according to the particular application. The amount administeredwill vary depending on the particular IC₅₀ value of the compound usedand the judgment of the attending clinician taking into considerationfactors such as health, weight, and age. In combinational applicationsin which the compound is not the sole active ingredient, it may bepossible to administer lesser amounts of compound and still havetherapeutic or prophylactic effect.

Preferably, the pharmaceutical preparation is in unit dosage form. Insuch form, the preparation is subdivided into unit doses containingappropriate quantities of the active component, e.g., an effectiveamount to achieve the desired purpose. The actual dosage employed may bevaried depending upon the requirements of the patient and the severityof the condition being treated. Determination of the proper dosage for aparticular situation is within the skill of the art. Generally,treatment is initiated with smaller dosages which are less than theoptimum dose of the compound. Thereafter, the dosage is increased bysmall amounts until the optimum effect under the circumstances isreached. For convenience, the total daily dosage may be divided andadministered in portions during the day if desired. The amount andfrequency of administration of the compounds and compositions of thepresent invention used in the methods of the present invention, and ifapplicable other chemotherapeutic agents and/or radiation therapy, willbe regulated according to the judgment of the attending clinician(physician) considering such factors as age, condition and size of thepatient as well as severity of the disease being treated.

The chemotherapeutic agent and/or radiation therapy can be administeredaccording to therapeutic protocols well known in the art. It will beapparent to those skilled in the art that the administration of thechemotherapeutic agent and/or radiation therapy can be varied dependingon the disease being treated and the known effects of thechemotherapeutic agent and/or radiation therapy on that disease. Also,in accordance with the knowledge of the skilled clinician, thetherapeutic protocols (e.g., dosage amounts and times of administration)can be varied in view of the observed effects of the administeredtherapeutic agents (i.e., antineoplastic agent or radiation) on thepatient, and in view of the observed responses of the disease to theadministered therapeutic agents.

Also, in general, the compounds of the invention need not beadministered in the same pharmaceutical composition as achemotherapeutic agent, and may, because of different physical andchemical characteristics, be administered by a different route. Forexample, the compounds/compositions may be administered orally togenerate and maintain good blood levels thereof, while thechemotherapeutic agent may be administered intravenously. Thedetermination of the mode of administration and the advisability ofadministration, where possible, in the same pharmaceutical composition,is well within the knowledge of the skilled clinician. The initialadministration can be made according to established protocols known inthe art, and then, based upon the observed effects, the dosage, modes ofadministration and times of administration can be modified by theskilled clinician.

The particular choice of compound (and where appropriate,chemotherapeutic agent and/or radiation) will depend upon the diagnosisof the attending physicians and their judgment of the condition of thepatient and the appropriate treatment protocol.

The compounds/compositions of the invention (and where appropriatechemotherapeutic agent and/or radiation) may be administeredconcurrently (e.g., simultaneously, essentially simultaneously or withinthe same treatment protocol) or sequentially, depending upon the natureof the proliferative disease, the condition of the patient, and theactual choice of chemotherapeutic agent and/or radiation to beadministered in conjunction (i.e., within a single treatment protocol)with the compound/composition. In combinational applications and uses,the compound/composition and the chemotherapeutic agent and/or radiationneed not be administered simultaneously or essentially simultaneously,and the initial order of administration of the compound/composition, andthe chemotherapeutic agent and/or radiation, may not be important. Thus,the compounds/compositions of the invention may be administered firstfollowed by the administration of the chemotherapeutic agent and/orradiation; or the chemotherapeutic agent and/or radiation may beadministered first followed by the administration of thecompounds/compositions of the invention. This alternate administrationmay be repeated during a single treatment protocol. The determination ofthe order of administration, and the number of repetitions ofadministration of each therapeutic agent during a treatment protocol, iswell within the knowledge of the skilled physician after evaluation ofthe disease being treated and the condition of the patient. For example,the chemotherapeutic agent and/or radiation may be administered first,especially if it is a cytotoxic agent, and then the treatment continuedwith the administration of the compounds/compositions of the inventionfollowed, where determined advantageous, by the administration of thechemotherapeutic agent and/or radiation, and so on until the treatmentprotocol is complete.

Thus, in accordance with experience and knowledge, the practicingphysician can modify each protocol for the administration of acompound/composition for treatment according to the individual patient'sneeds, as the treatment proceeds. The attending clinician, in judgingwhether treatment is effective at the dosage administered, will considerthe general well-being of the patient as well as more definite signssuch as relief of disease-related symptoms, inhibition of tumor growth,actual shrinkage of the tumor, or inhibition of metastasis. Size of thetumor can be measured by standard methods such as radiological studies,e.g., CAT or MRI scan, and successive measurements can be used to judgewhether or not growth of the tumor has been retarded or even reversed.Relief of disease-related symptoms such as pain, and improvement inoverall condition can also be used to help judge effectiveness oftreatment.

Assays for Determining HSP90 Binding and Downstream Effect

A variety of in vitro and in vivo assays are available to test theeffect of the alkynyl pyrrolo[2,3-d]pyrimidine compounds of theinvention on HSP90. HSP90 competitive binding assays and functionalassays can be performed as known in the art by substituting in thecompounds of the invention. Chiosis et al. Chemistry & Biology 2001, (S,289-299, describe some of the known ways in which this can be done. Forexample, competition binding assays using, e.g., geldanamycin or 17-AAGas a competitive binding inhibitor of HSP90 can be used to determinerelative HSP90 affinity of the compounds of the invention byimmobilizing the compound of interest or other competitive inhibitor ona gel or solid matrix, preincubating HSP90 with the other inhibitor,passing the preincubated mix over the gel or matrix, and then measuringthe amount of HSP90 that retains or does not retain on the gel ormatrix.

Downstream effects can also be evaluated based on the known effect ofHSP90 inhibition on function and stability of various steroid receptorsand signaling proteins including, e.g., Raf1 and Her2. Compounds of thepresent invention induce dose-dependent degradation of these molecules,which can be measured using standard techniques. Inhibition of HSP90also results in up-regulation of HSP90 and related chaperone proteinsthat can similarly be measured. Antiproliferative activity on variouscancer cell lines can also be measured, as can morphological andfunctional differentiation related to HSP90 inhibition.

Many different types of methods are known in the art for determiningprotein concentrations and measuring or predicting the level of proteinswithin cells and in fluid samples. Indirect techniques include nucleicacid hybridization and amplification using, e.g., polymerase chainreaction (PCR). These techniques are known to the person of skill andare discussed, e.g., in Sambrook, Fritsch & Maniatis Molecular Cloning:A Laboratory Manual, 2nd ed., Cold Spring Harbor Laboratory, Cold SpringHarbor, N.Y., 1989; Ausubel, et al. Current Protocols in MolecularBiology, John Wiley & Sons, NY, 1994, and, as specifically applied tothe quantification, detection, and relative activity of HER2/Neu inpatient samples, e.g., in U.S. Pat. Nos. 4,699,877, 4,918,162,4,968,603, and 5,846,749. A brief discussion of two generic techniquesthat can be used follows.

The determination of whether cells overexpress or contain elevatedlevels of Her2 can be determined using well known antibody techniquessuch as immunoblotting, radioimmunoassays, western blotting,immunoprecipitation, enzyme-linked immunosorbant assays (ELISA), andderivative techniques that make use of antibodies directed against Her2.As an example, Her2 expression in breast cancer cells can be determinedwith the use of an immunohistochemical assay, such as the Dako Hercep™test (Dako Corp., Carpinteria, Calif.). The Hercep™ test is an antibodystaining assay designed to detect Her2 overexpression in tumor tissuespecimens. This particular assay grades Her2 expression into fourlevels: 0, 1, 2, and 3, with level 3 representing the highest level ofHer2 expression. Accurate quantitation can be enhanced by employing anAutomated Cellular Imaging System (ACIS) as described, e.g., by Press,M. et al. Modern Pathology 2000, 13, 225A.

Antibodies, polyclonal or monoclonal, can be purchased from a variety ofcommercial suppliers, or may be manufactured using well-known methods,e.g., as described in Harlow et al. Antibodies: A Laboratory Manual, 2nded; Cold Spring Harbor Laboratory, Cold Spring Harbor, N.Y., 1988. Her2overexpression can also be determined at the nucleic acid level sincethere is a reported high correlation between overexpression of the Her2protein and amplification of the gene that codes for it. One way to testthis is by using RT-PCR. The genomic and cDNA sequences for Her2 areknown. Specific DNA primers can be generated using standard, well-knowntechniques, and can then be used to amplify template already present inthe cell. An example of this is described in Kurokawa, H. et al. CancerRes. 2000, 60, 5887-5894. PCR can be standardized such that quantitativedifferences are observed as between normal and abnormal cells, e.g.,cancerous and noncancerous cells. Well known methods employing, e.g.,densitometry, can be used to quantitate and/or compare nucleic acidlevels amplified using PCR. Similarly, fluorescent in situ hybridization(FISH) assays and other assays can be used, e.g., Northern and/orSouthern blotting. These rely on nucleic acid hybridization between theHer2 gene or mRNA and a corresponding nucleic acid probe that can bedesigned in the same or a similar way as for PCR primers, above. See,e.g., Mitchell M S, and Press M. F. Oncol., Suppl. 1999, 12, 108-116.For FISH, this nucleic acid probe can be conjugated to a fluorescentmolecule, e.g., fluorescein and/or rhodamine, that preferably does notinterfere with hybridization, and which fluorescence can later bemeasured following hybridization. See, e.g., Kurokawa, H et al, CancerRes. 2000, 60, 5887-5894 (describing a specific nucleic acid probehaving sequence 5′-FAM -NucleicAcid-TAMRA-p-3′ sequence). ACIS-basedapproaches as described above can be employed to make the assay morequantitative (de la Torre-Bueno, J., et al. Modern Pathology 2000, 13,221A).

Immuno and nucleic acid detection can also be directed against proteinsother than HSP90 and HER2, which proteins are nevertheless affected inresponse to HSP90 inhibition.

The following examples are offered by way of illustration only and arenot intended to be limiting of the full scope and spirit of theinvention.

EXAMPLES Materials and Methods

The chemical reagents used to create the novel products of the inventionbelow are all available commercially, e.g., from Aldrich Chemical Co.,Milwaukee, Wis., USA. Otherwise their preparation is facile and known toone of ordinary skill in the art, or it is referenced or describedherein.

The final compounds were usually purified by preparative TLC (silica gel60 Å, Whatman Partisil PK6F) or flash chromatography (silica gel 60 Å,EMD Chemicals) using EtOAc/hexane or MeOH/CH₂Cl₂ as eluents. Rf's weremeasured using silica gel TLC plates (silica gel 60 Å, EMED Chemicals).Analytical HPLC chromatograms were obtained using a C18 column (AgilentZorbax 300SB-C18; 5 microns; 4.6 mm×150 mm). A gradient was appliedbetween solvent A (0.1% TFA in H₂O) and solvent B (0.5% TFA in CH₃CN)increasing the proportion of A linearly from 5% (t=0) to 100% (t=7.00min), with a constant flow rate of 1 mL/min. The samples were diluted totypically 0.1-1 mg/ml in MeOH or CH₃CN and the injection volumes weretypically 10 μL. The column was not heated, and UV detection waseffected at 254 nm. ¹H-NMR spectra were recorded on a Bruker Avance 400MHz spectrometer.

The chemical names were generated using the Beilstein Autonom 2.1software.

General Procedures

General Procedure A: Sonogashira Coupling.

A mixture of the appropriate 5-iodo-pyrrolo[2,3-d]pyrimidine, alkyne(2-5 equiv.), Et₃N (2-5 equiv.), Pd(PPh₃)₄ (0.01-0.05 equiv.), and CuI(0.05-0.30 equiv.) in DCM (5 mL/mmol of starting iodide) was heated toreflux for 0.5-3 h. The reaction mixture was washed with sat. sq. NaHCO₃and brine, dried (Na₂SO₄), concentrated. Flash chromatography(EtOAc/DCM/Et₃N 15:74:1, gradually adding MeOH (0-7 vol %)) gave thedesired 5-alkynyl-pyrrolo[2,3-d]pyrimidine in typically 20-70% yield.

General Procedure B: ZnCl₂-Mediated Deprotection.

A suspension of 2-(pivaloylamino)-pyrrolo[2,3-d]pyrimidine and ZnCl₂(3-20 equiv.) in wet EtOH (5 vol % water) was heated to 80 AC, andmonitored by HPLC. When the reaction reached completion, DCM was added,and the organic layer was washed with sat. aq. NaHCO₃ and brine. Drying(Na₂SO₄) and concentration afforded the desired2-amino-pyrrolo[2,3-d]pyrimidine.

Example 13-[2-Amino-4-chloro-7-(4-methoxy-3,5-dimethyl-pyridin-2-ylmethyl)-7H-pyrrolo[2,3-d]pyrimidin-5-yl]-prop-2-yn-1-ol

See Shih, C. et al. Heterocycles, 1993, 35, 825 and U.S. Pat. No.5,196,424.

Step 1. 2-Amino-3,7-dihydro-pyrrolo[2,3-d]pyrimidin-4-one

A mixture of 2,4-diamino-6-hydroxypyrimidine (300 g, 2.37 mol),chloroacetaldehyde (50% aq. solution, 382 g, 2.43 mol, 303 mL, 1.02eq.), sodium acetate (195 g, 2.37 mol), DMF (2.5 L), and water (360 mL)was stirred mechanically at rt for 2 days. The resulting solid wascollected by filtration, and washed with water (50 mL×3). The motherliquor was concentrated to give additional material which was washedwith water (50 mL×3). The combined solid materials were recrystallizedfrom MeOH to give the title compound as a white powder (186 g, 52%yield, HPLC purity: 100%). t_(R): 2.21 min. ¹H-NMR (DMSO-d₆) δ 11.00(br. s, 1H), 10.33 (br. s, 1H), 6.63 (q, 1H), 6.21 (q, 1H), 6.12 (br. s,2H).

Step 2.2,2-Dimethyl-N-(4-oxo-4,7-dihydro-3H-pyrrolo[2,3-d]pyrimidin-2-yl)-propionamide

A solution of 2-amino-3,7-dihydro-pyrrolo[2,3-d]pyrimidin-4-one (186 g,1.23 mol) in pyridine (2 L) was treated with trimethylacetyl chloride(475 g, 3.94 mol, 485 mL, 3.2 eq) at 90° C. for 2 h, to give a mixtureof N(2)-monoacylated and N(2), N(7)-bisacylated material. The solventwas evaporated and the residue was taken up in aqueous ammonia (37% NH₃,310 mL) and MeOH (2 L), and stirred at rt for 30 min, to selectivelycleave the N(7)-pivaloyl group. The solid was collected by filtration,washed with water (500 mL×5), and dried on high vacuum to give the titlecompound as a solid (193 g). Concentration of the mother liquor providedadditional solid material, which was collected, washed with water (50mL×5), and dried (77 g). The combined yield was 93% (HPLC purity 98.6%).t_(R): 4.57 min. ¹H-NMR (DMSO-d₆) δ 12.00, (br. s, 1H), 7.40 (br. s,1H), 6.96 (q, 1H), 6.40 (q, 1H), 1.24 (s, 9H).

Step 3.N-(4-Chloro-7H-pyrrolo[2,3-d]pyrimidin-2-yl)-2,2-dimethyl-propionamide

A mixture of2,2-dimethyl-N-(4-oxo-4,7-dihydro-3H-pyrrolo[2,3-d]pyrimidin-2-yl)-propionamide(210 g, 0.90 mol), POCl₃ (828 g, 5.40 mol, 503 mL, 6.0 eq),benzyltriethylammonium chloride (411 g, 1.80 mol), N,N-dimethylaniline(220 g, 231 mL, 1.80 mol), and acetonitrile (2.0 L) was heated to refluxfor 40-60 min, monitoring with HPLC. The solvent was evaporated on arotary evaporator, and the residue was carefully (caution: exothermicand corrosive) and slowly added to ice water (16 L). The pH was adjustedto 7 with solid NaOH, and the resulting precipitate was collected byfiltration. Drying afforded the title compound (159 g, 70% yield 70%,HPLC purity 100%). t_(R): 5.37 min. ¹H-NMR (DMSO-d6) δ 12.35 (br. s,1H), 10.06 (br. S, 1H), 7.54 (q, 1H), 6.52 (q, 1H), 1.25 (s, 9H).

Step 4.N-(4-Chloro-5-iodo-7H-pyrrolo[2,3-d]pyrimidin-2-yl)-2,2-dimethyl-propionamide

A solution ofN-(4-chloro-7H-pyrolo[2,3-d]pyrimidin-2-yl)-2,2-dimethyl-propionamide(101 g, 0.40 mol) in anhydrous THF (2 L) was treated withN-4-iodosuccinimide (98.9 g, 0.44 mol, 1.1 eq.) under N₂ atmosphere atrt for 40 min. The solvent was evaporated, and the residue was taken upin CH₂Cl₂ (1.5 L), and washed with Na₂SO₃ (500 mL×3) and brine (300mL×3). Evaporated and recrystallizion from MeOH gave the title productas a white powder (122 g, 81% yield, HPLC purity: 98.2%). t_(R): 6.19min. ¹H-NMR (DMSO-d₆) δ 12.65 (br.s, 1H), 10.11 (br. s, 1H), 7.76 (d,1H), 1.24 (s, 9H).

Step 5.N-[4-Chloro-5-iodo-7-(4-methoxy-3,5-dimethyl-pyridin-2-ylmethyl)-7H-pyrrolo[2,3-d]pyrimidin-2-yl]-2,2-dimethyl-propionamide

A mixture ofN-(4-chloro-5-iodo-7H-pyrrolo[2,3-d]pyrimidin-2-yl)-2,2-dimethyl-propionamide(37.8 g, 0.1 mol), 2-chloromethyl-4-methoxy-3,5-dimethyl-pyridinehydrochloride (23.1 g, 0.104 mol), finely powdered K₂CO₃, (41.5 g, 0.3mol, 3.0 eq.), and anhydrous DMF (200 mL) was stirred at rt overnight.The solvent was evaporated, and the residue taken up in CH₂Cl₂ (500mL),washed with brine (200 mL×3), evaporated, and recrystallized from MeOHto give the title product as a white powder (42.0 g, 80% yield; HPLCpurity 98%). t_(R): 6.49 min. ¹H-NMR (DMSO-d₆) δ 10.16 (br. s, 1H), 8.04(s, 1H), 7.72 (s, 1H), 5.46 (s, 2H), 3.73 (s, 3H), 2.33 (s, 3H), 2.15(s, 3H), 1.21 (s, 9H).

Step 6.4-Chloro-5-iodo-7-(4-methoxy-3,5-dimethyl-pyridin-2-ylmethyl)-7H-pyrrolo[2,3-d]pyrimidin-2-ylamine

A mixture ofN-[4-chloro-5-iodo-7-(4-methoxy-3,5-dimethyl-pyridin-2-ylmethyl)-7H-pyrrolo[2,3-d]pyrimidin-2-yl]-2,2-dimethyl-propionamide(1.0 g, 1.89 mmol), ZnCl₂ (1.29 g, 9.47 mol, 5.0 eq.), and EtOH/H₂Osolution (25 mL, volumetric ratio 100:5) was stirred at 80° C.overnight. The reaction mixture poured into water, and the solid wascollected by filtration, washed with water (10 mL×3), and recrystallizedfrom MeOH to give the title product (0.67 g, 85% yield; HPLC purity98%). t_(R): 5.39 min. ¹H-NMR (DMSO-d₆) δ 8.07 (s, 1H), 7.28 (s, 1H),6.75 (br. s, 2H), 5.29 (s, 2H), 3.73 (s, 3H), 2.26 (s, 3H), 2.17 (s,3H).

Step 7.3-[2-Amino-4-chloro-7-(4-methoxy-3,5-dimetlzyl-pyridin-2-ylmethyl)-7H-pyrrolo[2,3-d]pyrimidin-5-yl]-prop-2-yn-1-ol

The title compound was prepared by Sonogashira coupling of4-Chloro-5-iodo-7-(4-methoxy-3,5-dimethyl-pyridin-2-ylmethyl)-7H-pyrrolo[2,3-d]pyrimidin-2-ylamaine(see example 1) with propargylic alcohol according to the GeneralProcedure A. t_(R): 4.42 min. ¹H-NMR (DMSO-d₆) δ 8.04 (s, 1H), 7.32 (s,1H), 6.71 (br. s, 2H), 5.27 (s, 2H), 5.22 (t, 1H), 4.26 (d, 1H), 3.71(s, 3H), 2.24 (s, 3H), 2.14 (s, 3H).

Example 24-[2-Amino-4-chloro-7-(4-methoxy-3,5-dimethyl-pyridin-2-ylmethyl)-7H-pyrrolo[2,3-d]pyrimidin-5-yl]-but-3-yn-1-ol

The title compound was prepared by Sonogashira coupling of4-chloro-5-iodo-7-(4-methoxy-3,5-dimethyl-pyridin-2-ylmethyl)-7H-pyrrolo[2,3-d]pyrimidin-2-ylamine(see Example 1) with 3-butyn-1-ol according to the General Procedure A.t_(R): 4.54 min. ¹H-NMR (DMSO-d₆) δ 8.04 (s, 1H), 7.22 (s, 1H), 6.69(br.s., 2H), 5.25 (s, 2H), 4.82 (d, 1H), 3.70 (s, 3H), 3.55 (q, 2H),2.49 (t, 2H), 2.23 (s, 3H), 2.14 (s, 3H).

Example 35-[2-Amino-4-chloro-7-(4-methoxy-3,5-dimethyl-pyridin-2-ylmethyl)-7H-pyrrolo[2,3-d]pyrimidin-5-yl]-pent-4-yn-1-ol

The title compound was prepared by Sonogashira coupling of4-chloro-5-iodo-7-(4-methoxy-3,5-dimethyl-pyridin-2-ylmethyl)-7H-pyrrolo[2,3-d]pyrimidin-2-ylamine(see Example 1) with 4-pentyn-1-ol according to the General Procedure A.t_(R): 4.71 min. ¹H-NMR (DMSO-d₆) δ 8.04 (s, 1H), 7.22 (s, 1H), 6.69(br. s, 2H), 5.25 (s, 2H), 4.48 (d, 1H), 3.71 (s, 3H), 3.50 (q, 2H),2.42 (t, 2H), 2.23 (s, 3H), 2.14 (s, 3H), 1.65 (5, 2H).

Example 44-Chloro-5-(3-diisobutylamino-prop-1-ynyl)-7-(4-methoxy-3,5-dimethyl-pyridin-2-ylmethyl)-7H-pyrrolo[2,3-d]pyrimidin-2-ylamine

Step 1.N-[4-Chloro-5-(3-hydroxy-prop-1-ynyl)-7-(4-methoxy-3,5-dimethyl-pyridin-2-ylmethyl)-7H-pyrrolo[2,3-d]pyrimidin-2-yl]-2,2-dimethyl-propionamide

The title compound was prepared by Sonogashira coupling ofN-[4-chloro-5-iodo-7-(4-methoxy-3,5-dimethyl-pyridin-2-ylmeth)-7H-pyrrolo[2,3-d]pyrimidin-2-yl]-2,2-dimethyl-propionamide(see Example 1) with propargylic alcohol according to the GeneralProcedure A. t_(R): 5.58 min. ¹H-NMR (DMSO-d₆): δ 10.15 (br. s, 1H),8.04 (s, 1H), 7.80 (s, 1H), 5.48 (s, 2H), 5.31 (t, 1H), 4.33 (d, 2H),3.74 (s, 3H), 2.33 (s, 3H), 2.15 (s, 3H), 1.21 (s, 9H).

Step 2.N-[4-Chloro-5-(3-diisobutylamino-prop-1-ynyl)-7-(4-methoxy-3,5-dimethyl-pyridin-2-ylmethyl)-7H-pyrrolo[2,3-d]pyrimidin-2-yl]-2,2-dimethyl-propionamide

A solution ofN-[4-chloro-5-(3-hydroxy-prop-1-ynyl)-7-(4-methoxy-3,5-dimethyl-pyridin-2-ylmethyl)-7H-pyrrolo[2,3-d]pyrimidin-2-yl]-2,2-dimethyl-propionamide(9.0 g, 19.7 mmol) and Et₃N (2 mL) in DCM (50 mL) was treated with MsCl(15.3 mL, 19.7 mmol) at 0° C. for 20 min and evaporated to give a yellowsolid, containing mostly the desired mesylate (10.7 g). An aliquot ofthis solid (106.8 mg, 0.2 mmol) was treated with diisobutylamine (25.9mg, 24.8 μL) in DCM (2 mL) at 0° C. for 10 h, and then at 25° C. for 3days. Evaporation and preparative plate chromatography gave the titlecompound.

Step 3.4-Chloro-5-(3-diisobutylamino-prop-1-ynyl)-7-(4-methoxy-3,5-dimethyl-pyridin-2-ylmethyl)-7H-pyrrolo[2,3-d]pyrimidin-2-ylamine

The title compound was prepared by cleaving the pivaloyl protectinggroup ofN-[4-chloro-5-(3-diisobutylamino-prop-1-ynyl)-7-(4-methoxy-3,5-dimethyl-pyridin-2-ylmethyl)-7H-pyrrolo[2,3-d]pyrimidin-2-yl]-2,2-dimethyl-propionamidewith ZnCl₂ according to the General Procedure B. t_(R): 5.23 min. ¹H-NMR(CDCl₃) δ 8.23 (s, 1H), 7.08 (s, 1H), 5.31 (s, 2H), 4.99 (s, 1H), 3.76(s, 3H), 3.56 (s, 2H), 2.29 (s, 31H), 2.27 (d, 4H), 2.21 (s, 31H), 1.74(m, 2H), 0.90 (d, 12H).

Example 54-Chloro-5-(3-diisopropylamino-prop-1-ynyl)-7-(4-methoxy-3,5-dimethyl-pyridin-2-ylmethyl)-7H-pyrrolo[2,3-d]pyrimidin-2-ylamine

Step 1.N-[4-Chloro-5-(3-diisopropylamino-prop-1-ynyl)-7-(4-methoxy-3,5-dimethyl-pyridin-2-ylmethyl)-7H-pyrrolo[2,3-d]pyrimidin-2-yl]-2,2-dimethyl-propionamide

A solution ofN-[4-chloro-5-(3-hydroxy-prop-1-ynyl)-7-(4-methoxy-3,5-dimethyl-pyridin-2-ylmethyl)-7H-pyrrolo[2,3-d]pyrimidin-2-yl]-2,2-dimethyl-propionamide(9.0 g, 19.7 mmol) and Et₃N (2 mL) in DCM (50 mL) was treated with MsCl(15.3 mL, 19.7 mmol) at 0° C. for 20 min and evaporated to give a yellowsolid, containing mostly the desired mesylate (10.7 g). An aliquot ofthis solid (106.8 mg, 0.2 mmol) was treated with diisopropylamine (20.3mg, 28.1 μL) in DCM (2 mL) at 0° C. for 10 h, and then at 25° C. for 3days. Evaporation and preparative plate chromatography gave the titlecompound.

Step 2.4-Chloro-5-(3-diisopropylamino-prop-1-ynyl)-7-(4-methoxy-3,5-dimethyl-pyridin-2-ylmethyl)-7H-pyrrolo[2,3-d]pyrimidin-2-ylamine

The title compound was prepared by cleaving the pivaloyl protectinggroup ofN-[4-chloro-5-(3-diisopropylamino-prop-1-ynyl)-7-(4-methoxy-3,5-dimethyl-pyridin-2-ylmethyl)-7H-pyrrolo[2,3-d]pyrimidin-2-yl]-2,2-dimethyl-propionamidewith ZnCl₂ according to the General Procedure B. t_(R): 4.71 min. ¹H-NMR(CDCl₃) δ 8.24 (s, 1H), 7.05 (s, 1H), 5.30 (s, 2H), 4.96 (s, 1H), 3.76(s, 3H), 3.68 (s, 2H), 3.27 (7, 2H), 2.27 (s, 2H), 2.20 (s, 2H), 1.15(d, 12H).

Example 64-Chloro-7-(4-methoxy-3,5-dimethyl-pyridin-2-ylmethyl)-5-pyridin-2-ylethynyl-7H-pyrrolo[2,3-d]pyrimidin-2-ylamine

The title compound was prepared by Sonogashira coupling of4-chloro-5-iodo-7-(4-methoxy-3,5-dimethyl-pyridin-2-ylmethyl)-7H-pyrrolo[2,3-d]pyrimidin-2-ylamine(see, Example 1) with 2-ethynylpyridine according to the GeneralProcedure A. t_(R): 4.65 min. ¹H-NMR (DMSO-d₆) δ 8.59 (s, 1H), 8.07 (s,1H), 7.82 (td, 1H), 7.62 (s, 1H), 7.56 (d, 1H), 7.36 (td, 1H), 6.84 (br.S, 2H), 5.36 (s, 2H), 3.74 (s, 3H), 2.28 (s, 3H), 2.17 (s, 3H).

Example 76-[2-Amino-4-chloro-7-(4-methoxy-3,5-dimethyl-pyridin-2-ylmethyl)-7H-pyrrolo[2,3-d]pyrimidin-5-yl]-hex-5-yn-1-ol

The title compound was prepared by Sonogashira coupling of4-chloro-5-iodo-7-(4-methoxy-3,5-dimethyl-pyridin-2-ylmethyl)-7H-pyrrolo[2,3-d]pyrimidin-2-ylamine(see Example 1) with 5-hexyn-1-ol according to the General Procedure A.t_(R): 4.92 min. ¹H-NMR (DMSO-d₆) δ 8.04 (s, 1H), 7.22 (s, 1H), 6.69(br. s, 2H), 5.25 (s, 2H), 4.39 (t, 1H), 3.70 (s, 3H), 3.40 (q, 2H),2.39 (t, 2H), 2.23 (s, 3H), 2.14 (s, 3H), 1.55 (m, 4H).

Example 84-Chloro-7-(4-methoxy-3,5-dimethyl-pyridin-2-ylmethyl)-5-trimethylsilanylethynyl-7H-pyrrolo[2,3-d]pyrimidin-2-ylamine

Step 1:N-[4-Chloro-7-(4-methoxy-3,5-dimethyl-pyridin-2-ylmethyl)-5-trimethylsilanylethynyl-7H-pyrrolo[2,3-d]pyrimidin-2-yl]-2,2-dimethyl-propionamide

The title compound was prepared by Sonogashira coupling ofN-[4-chloro-5-iodo-7-(4-methoxy-3,5-dimethyl-pyridin-2-ylmethyl)-7H-pyrrolo[2,3-d]pyrimidin-2-yl]-2,2-dimethyl-propionamide(see Example 1) with ethynyl-trimethylsilane according to the GeneralProcedure A. t_(R)=7.48 min. ¹H NMR (CDCl₃) δ 8.18 (s, 2H), 7.38 (s,1H), 5.49 (s, 2H), 3.74 (s, 3H), 2.24 (s, 6H), 1.35 (s, 9H), 0.24 (s,9H).

Step 2:4-Chloro-7-(4-methoxy-3,5-dimethyl-pyridin-2-ylmethyl)-5-trimethylsilanylethynyl-7H-pyrrolo[2,3-d]pyrimidin-2-ylamine

The title compound was prepared by cleaving the pivaloyl protectinggroup ofN-[4-chloro-7-(4-methoxy-3,5-dimethyl-pyridin-2-ylmethyl)-5-trimethylsilanylethynyl-7H-pyrrolo[2,3-d]pyrimidin-2-yl]-2,2-dimethyl-propionamide with ZnCl₂ according to the GeneralProcedure B. t_(R)=6.56 min. ¹H NMR (CDCl₃) δ 8.22 (s, 1H), 7.11 (s,1H), 5.30 (s, 2H), 5.09 (s, 2H), 3.75 (s, 3H), 2.26 (s, 3H), 2.18 (s,3H), 0.24 (s, 9H).

Example 95-[2-Amino-4-chloro-7-(4-methoxy-3,5-dimethyl-pyridin-2-ylmethyl)-7H-pyrrolo[2,3-d]pyrimidin-5-yl]-pent-4-ynoicacid

The title compound was prepared by Sonogashira coupling of4-Chloro-5-iodo-7-(4-methoxy-3,5-dimethyl-pyridin-2-ylmethyl)-7H-pyrrolo[2,3-d]pyrimidin-2-ylamine(see Example 1) with 4-pentynoic acid according to the General ProcedureA. t_(R)=4.73 min. ¹H NMR (DMSO-d) δ 12.2 (s, 1H), 8.15 (s, 1H), 7.28(s, 1H), 6.80 (s, 2H), 5.33 (s, 2H), 3.73 (s, 3H), 3.31 (br. s, 2H),2362 (br. t, 2H), 2.27 (s, 3H), 2.17 (s, 3H).

Example 105-[2-Amino-4-chloro-7-(4-methoxy-3,5-dimethyl-pyridin-2-ylmethyl)-7H-pyrrolo[2,3-d]pyrimidin-5-yl]-pent-4-ynoicacid diethylamide

A mixture of5-[2-amino-4-chloro-7-(4-methoxy-3,5-dimethyl-pyridin-2-ylmethyl)-7H-pyrrolo[2,3-d]pyrimidin-5-yl]-pent-4-ynoic acid (see previous example, 25 mg),Et₂NH (100 μL), EDCI (87 mg), HOBt (50 mg), and DMF (1 mL) was stirredat rt for 16 h. Preparative plate chromatography (EtOAc/hexane 2:1) gavethe title compound. t_(R)=5.33 min. ¹H NMR (CDCl₃) δ 8.23 (s, 1H), 7.01(s, 1H), 5.32 (s, 2H), 4.98 (s, 2H), 3.76 (s, 3H), 3.42 (q, 2H), 3.33(q, 2H), 2.78 (dd, 2H), 2.64 (dd, 2H), 2.27 (s, 3H), 2.18 (s, 3H), 1.20(t, 3H), 1.13 (t, 3H).

Example 115-[2-Amino-4-chloro-7-(4-methoxy-3,5-dimethyl-pyridin-2-ylmethyl)-7H-pyrrolo[2,3-d]pyrimidin-5-yl]-1-(4-methyl-piperazin-1-yl)-pent-4-yn-1-one

Step 1: 1-(4-Methyl-piperazin-1-yl)-pent-4-yn-1-one

A solution of 4-pentynoic acid (519 mg, 5.29 mmol) and Et₃N (737 μL,5.29 mmol) in DCM (10 mL) was treated with ethyl chloroformate (504 μL,5.29 mmol) at rt for 15 min. Then N-methyl-1-piperazine (588 μL, 5.29mmol) was added, and stirring was prolonged for 45 min. Work-up (sat.aq. NaHCO₃) and concentration gave the title compound as a colorlessoil. ¹H NMR (CDCl₃) δ 3.63 (t, 2H), 3.48 (t, 2H), 2.55 (m, 4H), 2.38(quint., 41H), 2.30 (s, 3H).

Step 2:5-[2-Amino-4-chloro-7-(4-methoxy-3,5-dimethyl-pyridin-2-ylmethyl)-7H-pyrrolo[2,3-d]pyrimidin-5-yl]-1-(4-methyl-piperazin-1-yl)-pent-4-yn-1-one

The title compound was obtained by Sonogashira coupling of4-Chloro-5-iodo-7-(4-methoxy-3,5-dimethyl-pyridin-2-ylmethyl)-7H-pyrrolo[2,3-d]pyrimidin-2-ylamine(see Example 1) with 1-(4-Methyl-piperazin-1-yl)-pent-4-yn-1-oneaccording to the General Procedure A. t_(R)=4.16 min. ¹H NMR (CDCl₃) δ8.22 (s, 1H), 7.00 (s, 1H), 5.30 (s, 2H), 5.14 (s, 2H), 3.74 (s, 3H),3.64 (t, 2H), 3.50 (t, 2H), 2.76 (dd, 2H), 2.64 (dd, 2H), 2.29 (s, 3H),2.24 (s, 3H), 2.18 (s, 3H).

Example 125-[2-Amino-4-chloro-7-(4-methoxy-3,5-dimethyl-pyridin-2-ylmethyl)-7H-pyrrolo[2,3-d]pyrimidin-5-yl]-pent-4-ynoicacid amide

Step 1: Pent-4-ynoic acid amide

A solution of 4-pentynoic acid (517 mg, 5.29 mmol) and Et₃N (737 μL,5.29 mmol) in DCM (10 mL) was treated with ethyl chloroformate (504 μL,5.29 mmol) at rt for 15 min. Then NH₃ (7M in MeOH, 1 mL, 7 mmol) wasadded, and stirring was prolonged for 5 min. Work-up (sat. aq. NaHCO₃)and concentration gave the title compound as a colorless solid. ¹H NMR(CDCl₃) δ 6.03 (s, 1H), 5.86 (s, 1H), 2.55 (m, 2H), 2.46 (m, 2H), 2.02(t, 1H).

Step 2:5-[2-Amino-4-chloro-7-(4-methoxy-3,5-dimethyl-yl-pyridin-2-ylmethyl)-7H-pyrrolo[2,3-d]pyrimidin-5-yl]-pent-4-ynoicacid amide

The title compound was obtained by Sonogashira coupling of4-Chloro-5-iodo-7-(4-methoxy-3,5-dimethyl-pyridin-2-ylmethyl)-7H-pyrrolo[2,3-d]pyrimidin-2-ylamine(see Example 1) with pent-4-ynoic acid amide according to the generalprocedure A. t_(R)=4.35 min. ¹H NMR (DMSO-d₆) δ 8.07 (s, 1H), 7.36 (s,1H), 7.23 (s, 1H), 6.87 (s, 1H), 6.71 (s, 2H), 5.28 (s, 2H), 3.72 (s,3H), 2.59 (t, 2H), 2.34 (t, 2H), 2.25 (s, 3H), 2.16 (s, 3H).

Example 13{3-[2-Amino-4-chloro-7-(4-methoxy-3,5-dimethyl-pyridin-2-ylmethyl)-7H-pyrrolo[2,3-d]pyrimidin-5-yl]-prop-2-ynyl}-carbamic acid tert-butyl ester

Step 1: Prop-2-ynyl-carbamic acid tert-butyl ester

A solution of propargylamine (1 g, 18 mmol) in DCM (10 mL) was treatedwith (BOC)₂O (4.0 g, 18 mmol) at rt for 3 h. Evaporation gave the titlecompound as an oil. ¹H NMR (CDCl₃) δ 4.80 (s, 1H), 3.93 (s, 2H), 2.23(t, 1H), 1.47 (s, 9H).

Step 2:{3-[2-Amino-4-chloro-7-(4-methoxy-3,5-dimethyl-pyridin-2-ylmethyl)-7H-pyrrolo[2,3-d]pyrimidin-5-yl]-prop-2-ynyl}-carbamicacid tert-butyl ester

The title compound was obtained by Sonogashira coupling of4-Chloro-5-iodo-7-(4-methoxy-3,5-dimethyl-pyridin-2-ylmethyl)-7H-pyrrolo[2,3-d]pyrimidin-2-ylamine(see Example 1) with prop -2-ynyl-carbamic acid tert-butyl esteraccording to the General Procedure A. t_(R)=5.91 min. ¹H NMR (CDCl₃) δ8.20 (s, 1H), 7.05 (s, 1H), 5.36 (s, 2H), 5.31 (s, 2H), 4.82 (s, 1H),4.16 (s, 2H), 3.76 (s, 3H), 2.26 (s, 3H), 2.24 (s, 3H), 1.47 (s, 9H).

Example 145-(3-Amino-prop-1-ynyl)-4-chloro-7-(4-methoxy-3,5-dimethyl-pyridin-2-ylmethyl)-7H-pyrrolo[2,3-d]pyrimidin-2-ylamine

A solution of{3-[2-Amino-4-chloro-7-(4-methoxy-3,5-dimethyl-pyridin-2-ylmethyl)-7H-pyrrolo[2,3-d]pyrimidin-5-yl]-prop-2-ynyl}-carbamic acid tert-butyl ester (seeprevious example, 22 mg) in DCM (3 mL) was treated with TFA (0.6 mL) atrt for 15 min. Evaporation, work-up (DCM/sat. aq. NaHCO₃), drying(Na₂SO₄), and evaporation gave the title compound as an oil. t_(R)=3.85min. ¹H NMR (CDCl₃) δ 8.24 (s, 1H), 7.06 (s, 1H), 5.32 (s, 2H), 5.02 (s,2H), 3.75 (s, 3H), 2.27 (s, 3H), 2.20 (s, 3H).

Example 15{5-[2-Amino-4-chloro-7-(4-methoxy-3,5-dimethyl-pyridin-2-ylmethyl)-7H-pyrrolo[2,3-d]pyrimidin-5-yl]-pent-4-ynyl}-carbamic acid tert-butyl ester

Step 1: Pent-4-ynyl-carbamic acid tert-butyl ester

A solution of pent-4-ynylamine (480 mg; Li, Y. et al. J. Am. Chem. Soc.1996, 118, 9295) in DCM (5 mL) was treated with (BOC)₂O at rt for 15min. Evaporation gave the title compound as a colorless oil. ¹H NMR(CDCl₃) δ 4.68 (s, 1H), 3.24 (q, 2H), 2.25 (td, 2H), 1.97 (t, 1H), 1.70(quint., 2H), 1.45 (s, 9H).

Step 2:{5-[2-Amino-4-chloro-7-(4-methoxy-3,5-dimethyl-pyridin-2-ylmethyl)-7H-pyrrolo[2,3-d]pyrimidin-5-yl]-pent-4-ynyl}-carbamicacid tert-butyl ester

The title compound was obtained by Sonogashira coupling of4-Chloro-5-iodo-7-(4-methoxy-3,5-dimethyl-pyridin-2-ylmethyl)-7H-pyrrolo[2,3-d]pyrimidin-2-ylamine(see Example 1) with pent-4-ynyl-carbamic acid tert-butyl esteraccording to the General Procedure A. t_(R)=5.96 min. ¹H NMR (CDCl₃) δ8.23 (s, 1H), 7.02 (s, 1H), 5.30 (s, 2H), 5.07 (s, 2H), 4.80 (br. t,1H), 3.75 (s, 3H), 3.30 (q, 2H), 2.48 (t, 2H), 2.26 (s, 3H), 2.19 (s,3H), 1.80 (quint., 2H), 1.43 (s, 9H).

Example 165-(5-Amino-pent-1-ynyl)-4-chloro-7-(4-methoxy-3,5-dimethyl-pyridin-2-ylmethyl)-7H-pyrrolo[2,3-d]pyrimidin-2-ylamine

A solution of{5-[2-amino-4-chloro-7-(4-methoxy-3,5-dimethyl-pyridin-2-ylmethy)-7H-pyrrolo[2,3-d]pyrimidin-5-yl]-pent-4-ynyl}-carbamic acid tert-butyl ester (seeprevious example, 33 mg) in DCM (1.0 mL) was treated with TFA (0.2 mL)at rt for 10 min. The reaction mixture was concentrated, taken in water(1.5 mL), washed with EtOAc (1.5 mL), made alkaline with sat. aq. NH₄OH(0.5 mL), and back-extracted into EtOAc (25 mL). Washing (NH₄OH 1M),drying (Na₂SO₄) and concentration afforded the title compound.t_(R)=4.37 min. ¹H NMR (CDCl₃) δ 8.15 (s, 1H), 6.97 (s, 1H), 5.25 (s,2H), 3.72 (s, 3H), 2.94 (m, 2H), 2.48 (t, 2H), 2.23 (s, 3H), 2.16 (s,3H), 1.80 (quint., 2H).

Example 172-{5-[2-Amino-4-chloro-7-(4-methoxy-3,5-dimethyl-pyridin-2-ylmethyl)-7H-pyrrolo[2,3-d]pyrimidin-5-yl]-pent-4-ynyl}-isoindole-1,3-dione

The title compound was obtained by Sonogashira coupling of4-Chloro-5-iodo-7-(4-methoxy-3,5-dimethyl-pyridin-2-ylmethyl)-7H-pyrrolo[2,3-d]pyrimidin-2-ylamine(see Example 1) with 2-pent -4-ynyl-isoindole-1,3-dione (Li, Y. et al.J. Am. Chem. Soc. 1996, 118, 9295) according to the General Procedure A.t_(R)=6.08 min. ¹H NMR (CDCl₂;) δ 8.17 (s, 1H), 7.75 (dd, 2H), 7.58 (dd,2H), 6.85 (s, 1H), 5.22 (s, 2H), 3.81 (t, 2H), 3.72 (s, 3H), 2.46 (t,2H), 2.23 (s, 3H), 2.14 (s, 3H), 1.95 (quint., 2H).

Example 184-Chloro-7-(4-methoxy-3,5-dimethyl-pyridin-2-ylmethyl)-5-(4-morpholin-4-yl-but-1-ynyl)-7H-pyrrolo[2,3-d]pyrimidin-2-ylamine

Step 1: Methanesulfonic acid but-3-ynyl ester

A solution of 3-but-1-ynol (5.62 g, 80.2 mmol.) and Et₃N (14.5 mL, 104mmol) in DCM (40 mL) was treated with MsCl (7.48 mL, 96.2 mmol) at 0° C.for 10 min. Work-up (water; sat. aq. NaHCO₃), drying (Na₂SO₄), andconcentration gave the title compound as a pale orange oil. ¹H NMR(CDCl₃) δ 4.32 (t, 2H), 3.07 (s, 3H), 2.67 (td, 2H), 2.09 (t, 2H).

Step 2: 4-But-3-ynyl-morpholine

A mixture of methanesulfonic acid but-3-ynyl ester (10.1 g, 68 mmol) andmorpholine (12.5 mL, 143 mmol) was heated to 100° C. for 1 h. Themixture was diluted with Et₂O (50 mL) and filtered. The solid residuewas discarded, and the mother liquor extracted with 3N HCl. The aqueouslayer was basified with NaOH, and back-extracted into EtOAc. Drying(Na₂SO₄), concentration, and distillation (short-path, b.p.=130° C. at˜5 mm Hg) game the title compound as a colorless oil. ¹H NMR (CDCl₃) δ3.72 (t, 4H), 2.60 (t, 2H), 2.48 (t, 4H), 2.39 (td, 2H), 2.00 (t, 1H).

Step 3:4-Chloro-7-(4-methoxy-3,5-dimethyl-pyridin-2-ylmethyl)-5-(4-morpholin-4-yl-but-1-ynyl)-7H-pyrrolo[2,3-d]pyrimidin-2-ylamine

The title compound was obtained by Sonogashira coupling of4-Chloro-5-iodo-7-(4-methoxy-3,5-dimethyl-pyridin-2-ylmethyl)-7H-pyrrolo[2,3-d]pyrimidin-2-ylamine(see Example 1) with 4-but -3-ynyl-morpholine according to the GeneralProcedure A. t_(R)=4.12 min. ¹H NMR (CDCl₃) δ 8.33 (s, 1H), 6.96 (s,1H), 5.26 (s, 2H), 5.12 (s, 2H), 3.76 (s, 3H), 3.72 (t, 4H), 2.67 (m,2H), 2.60 (m, 2H), 2.52 (t, 4H), 2.25 (s, 3H), 2.20 (s, 3H).

Example 194-Chloro-7-(4-methoxy-3,5-dimethyl-pyridin-2-ylmethyl)-5-(5-morpholin-4-yl-pent-1-ynyl)-7H-pyrrolo[2,3-d]pyrimidin-2-ylamine

Step 1: 5-Iodo-pent-1-yne

A mixture of 5-chloro-pent-1-yne (5.0 mL, 47 mmol), NaI (9.7 g, 65 mmol)and acetone (20 mL) was heated to reflux for 15 h. Additional NaI (9.7g, 65 mmol) was added, and the refluxed was prolonged for 24 h. Afterfiltration and concentration, the residue was dissolved in hexane andwashed with water and aq. Na₂S₂O₃. Drying (Na₂SO₄) and concentrationafforded the title compound. ¹H NMR (CDCl₃) δ 3.33 (t, 2H), 2.36 (td,2H), 2.03 (quint., 2H), 2.01 (t, 1H).

Step 2: 4-Pent-4-ynyl-morpholine

A mixture of 5-iodo-pent-1-yne (2.40 g, 12.3 mmol) and morpholine (2.70g, 30.9 mmol) was heated to 80° C. for 15 min. The mixture was dilutedwith Et₂O and filtered. The solid residue was discarded, and the motherliquor was extracted with 3N HCl. The aqueous layer was made basic withNaOH, and back-extracted into EtOAc. Drying (Na₂SO₄), and concentrationgave the title compound as a pale orange oil. ¹H NMR (CDCl₃) δ 3.67 (t,4H), 2.39 (m, 6H), 2.21 (td, 2H), 1.93 (t, 1H), 1.67 (quint., 2H).

Step 3:4-Chloro-7-(4-methoxy-3,5-dimethyl-pyridin-2-ylmethyl)-5-(5-morpholin-4-yl-pent-1-ynyl)-7H-pyrrolo[2,3-d]pyrimidin-2-ylamine

The title compound was obtained by Sonogashira coupling of4-Chloro-5-iodo-7-(4-methoxy-3,5-dimethyl-pyridin-2-ylmethyl)-7H-pyrrolo[2,3-d]pyrimidin-2-ylamine(see Example 1) with 4-pent -4-ynyl-morpholine according to the generalprocedure A. t_(R)=4.26 min. ¹H NMR (CDCl₃) δ 8.21 (s, 1H), 7.00 (s,1H), 5.29 (s, 2H), 5.13 (s, 2H), 3.74 (s, 3H), 3.71 (t, 4H), 2.51-2.45(m, 8H), 2.25 (s, 3H), 2.18 (s, 3H), 1.78 (quint., 2H).

Example 205-(4-tert-Butylamino-but-1-ynyl)-4-chloro-7-(4-methoxy-3,5-dimethyl-pyridin-2-ylmethyl)-7H-pyrrolo[2,3-d]pyrimidin-2-ylamine

Step 1: tert-Butyl-but-3-ynyl-amine

A solution of toluene-4-sulfonic acid but-3-ynyl ester (1.57 g, 7.1mmol) in tert-BuNH₂ (3.54 g. 48 mmol) was heated to reflux for 18 h. Thereaction mixture was diluted with Et₂O (20 mL), filtered, andconcentrated to afford the title product. ¹H NMR (CDCl₃) δ 2.74 (t, 2H),2.38 (td, 2H), 2.00 (t, 1H), 1.12 (s, 9H).

Step 2:5-(4-tert-Butylamino-but-1-ynyl)-4-chloro-7-(4-methoxy-3,5-dimethyl-pyridin-2-ylmethyl)-7H-pyrrolo[2,3-d]pyrimidin-2-ylamine

The title compound was obtained by Sonogashira coupling of4-Chloro-5-iodo-7-(4-methoxy-3,5-dimethyl-pyridin-2-ylmethyl)-7H-pyrrolo[2,3-d]pyrimidin-2-ylamine(see Example 1) with tert-butyl-but -3-ynyl-amine according to theGeneral Procedure A. t_(R)=4.38 min. ¹H NMR (CDCl₃) δ 8.21 (s, 1H), 7.03(s, 1H), 5.29 (s, 2H), 5.10 (s, 2H), 3.74 (s, 3H), 2.81 (t, 2H), 2.62(t, 2H), 2.25 (s, 3H), 2.18 (s, 3H), 1.13 (s, 9H).

Example 215-(5-tert-Butylamino-pent-1-ynyl)-4-chloro-7-(4-methoxy-3,5-dimethyl-pyridin-2-ylmethyl)-7H-pyrrolo[2,3-d]pyrimidin-2-ylamine

Step 1: tert-Butyl-pent-4-ynyl-amine

A solution of 5-iodo-pent-1-yne (1.0 g, 5.2 mmol) in tert-BuNH₂ (2.0 g.27 mmol) was heated to reflux for 4 h, evaporated, diluted with Et₂O,filtered, and concentrated to afford the title compound. ¹H NMR (CDCl₃)δ 2.67 (t, 2H), 2.27 (td, 2H), 1.96 (t, 1H), 1.70 (quint., 2H), 1.11 (s,9H).

Step 2:5-(5-tert-Butylamino-pent-1-ynyl)-4-chloro-7-(4-methoxy-3,5-dimethyl-pyridin-2-ylmethyl)-7H-pyrrolo[2,3-d]pyrimidin-2-ylamine

The title compound was obtained by Sonogashira coupling of4-Chloro-5-iodo-7-(4-methoxy-3,5-dimethyl-pyridin-2-ylmethyl)-7H-pyrrolo[2,3-d]pyrimidin-2-ylamine(see Example 1) with tert-butyl-pent -4-ynyl-amine according to theGeneral Procedure A. t_(R)=4.51 min. ¹H NMR (CDCl₃) δ 8.23 (s, 1H), 7.02(s, 1H), 5.30 (s, 2H), 5.00 (s, 2H), 3.75 (s, 3H), 2.74 (t, 2H), 2.50(t, 2H), 2.27 (s, 3H), 2.19 (s, 3H), 1.76 (quint., 2H), 1.12 (s, 9H).

Example 22 Dimethylamino-acetic acid3-[2-amino-4-chloro-7-(4-methoxy-3,5-dimethyl-pyridin-2-ylmethyl)-7H-pyrrolo[2,3-d]pyrimidin-5-yl]-prop-2-ynylester

A solution of3-[2-amino-4-chloro-7-(4-methoxy-3,5-dimethyl-pyridin-2-ylmethyl)-7H-pyrrolo[2,3-d]pyrimidin-5-yl]-prop-2-yn-1-ol (see example 1, 177 mg, 0.48 mmol)in anhydrous pyridine (3 mL) was treated with N,N-dimethylamino-acetylchloride hydrochloride (113 mg, 0.72 mmol) at rt for 30 min. Addition oftoluene (10 mL) caused the formation of a sticky pellet. The toluenesolution was discarded, and the pellet was partitioned between water andDCM. The DCM layer was concentrated, and purified by flashchromatography (EtOAc/DCM/Et₃N 33:66:1, gradually adding MeOH (0-2%)) toafford the title compound. t_(R)=4.51 min. ¹H NMR (CDCl₃) δ 8.23 (s,1H), 7.14 (s, 1H), 5.31 (s, 2H), 5.00 (s, 2H), 4.98 (s, 2H), 3.76 (s,3H), 3.25 (s, 2H), 2.38 (s, 6H), 2.28 (s, 3H), 2.20 (s, 3H).

Example 235-(2-amino-4-chloro-7-((4-methoxy-3,5-dimethylpyridin-2-yl)methyl)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-methylpent-4-yn-2-ol

Sonogashira coupling of4-chloro-5-iodo-7-((4-methoxy-3,5-dimethylpyridin-2-yl)methyl)-7H-pyrrolo[2,3-d]pyrimidin-2-amine with 2-methylpent-4-yn-2-ol (H. Zhang et al.,Tetrahedron Lett. 1999, 40, 7851) according to the general procedure Agave the title compound, as a solid. Mp=163-165° C. HPLC Rt=4.98 min.

Example 244-chloro-7-((4-methoxy-3,5-dimethylpyridin-2-yl)methyl)-5-(5-(4-methylpiperazin-1-yl)pent-1-ynyl)-7H-pyrrolo[2,3-d]pyrimidin-2-amine

Step 1: 1-methyl-4-(pent-4-ynyl)piperazine

A mixture of pent-4-ynyl methanesulfonate (887 mg) and N-methylpiperazine (607 μL) was heated to 80° C. for 4.5 h, diluted with1,2-dichloroethane (5 mL) and heated to 70° C. for 18 h. Saturated aq.NaHCO₃ was added (10 mL) and the mixture was extracted with DCM (3×40mL), dried over Na₂SO₄, and concentrated to afford1-methyl-4-(pent-4-ynyl)piperazine (0.23 g).

Step 2

Sonogashira coupling of4-chloro-5-iodo-7-((4-methoxy-3,5-dimethylpyridin-2-yl)methyl)-7H-pyrrolo[2,3-d]pyrimidin-2-amine with 1-methyl-4-(pent-4-ynyl)piperazine,according to the general procedure A gave the title compound, as asolid. Mp=160.1-162.3° C. HPLC Rt=4.08 min.

Example 254-chloro-5-(5-(4-ethylpiperazin-1-yl)pent-1-ynyl)-7-((4-methoxy-3,5-dimethylpyridin-2-yl)methyl)-7H-pyrrolo[2,3-d]pyrimidin-2-amine

Step 1: 1-ethyl-4-(pent-4-ynyl)piperazine

A mixture of pent-4-ynyl methanesulfonate (849 mg) and N-methylpiperazine (665 μL) was heated to 80° C. for 3 h, diluted with1,2-dichloroethane (5 mL) and heated to 70° C. for 18 h. Saturated aq.NaHCO₃ was added (10 mL) and the mixture was extracted with DCM (1×40mL), dried over Na₂SO₄, and concentrated to afford 0.56 g of1-ethyl-4-(pent-4-ynyl)piperazine.

Step 2

Sonogashira coupling of4-chloro-5-iodo-7-((4-methoxy-3,5-dimethylpyridin-2-yl)methyl)-7H-pyrrolo[2,3-d]pyrimidin-2-amine with 1-ethyl-4-(pent-4-ynyl)piperazine,according to the general procedure A gave the title compound, as asolid. Mp=142.3-144.1° C. HPLC Rt=4.11 min.

Example 264-chloro-7-((4-methoxy-3,5-dimethylpyridin-2-yl)methyl)-5-(4-(4-methylpiperazin-1-yl)but-1-ynyl)-7H-pyrrolo[2,3-d]pyrimidin-2-amine

Step 1: 1-(but-3-ynyl)-4-methylpiperazine

A mixture of but-3-ynyl 4-methylbenzenesulfonate (972 mg) and N-methylpiperazine (482 μL) was heated to 80 C for 4.5 h, diluted with1,2-dichloroethane (5 mL) and heated to 70° C. for 18 h. Saturated aq.NaHCO₃ was added (10 mL) and the mixture was extracted with DCM (3×40mL), and concentrated to afford 0.70 g of1-(but-3-ynyl)-4-methylpiperazine as a 3:2 mixture of tosylate salt andfree base.

Step 2

Sonogashira coupling of4-chloro-5-iodo-7-((4-methoxy-3,5-dimethylpyridin-2-yl)methyl)-7H-pyrrolo[2,3-d]pyrimidin-2-amine with 1-(but-3-ynyl)-4-methylpiperazineaccording to the general procedure A gave the title compound, as an oil.HPLC Rt=4.05 min.

Example 274-(5-(2-amino-4-chloro-7-((4-methoxy-3,5-dimethylpyridin-2-yl)methyl)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)pent-4-ynyl)-N-methylpiperazine-1-carboxamide

Step 1: 1-(pent-4-ynyl)piperazine

A solution of pent-4-ynyl methanesulfonate (19.55 g) and piperazine(41.5 g) in EtOH (80 mL) was heated to reflux for 1 h, and evaporated.The residue was taken in NaOH 2M (70 mL) and extracted with Et₂O (70mL). Evaporation gave 1-(pent-4-ynyl)piperazine (4.4 g).

Step 2: 4-nitrophenyl 4-(pent-4-ynyl)piperazine-1-carboxylate

A solution of 1-(pent-4-ynyl)piperazine (2.2 g) and Et₃N (22 mL) in1,2-dichloroethane (22 mL) was treated with p-nitrophenyl chloroformate(2.9 g) at rt for 10 min. Work-up and silica gel chromatography(DCM:EtOAc 1:3→0:3) gave 4-nitrophenyl4-(pent-4-ynyl)piperazine-1-carboxylate as a yellow oil (2.96 g).

Step 3: N-methyl-4-(pent-4-ynyl)piperazine-1-carboxamide

A solution of 4-nitrophenyl 4-(pent-4-ynyl)piperazine-1-carboxylate(1.37 g) in THF (10 mL) was treated with 40% aq. MeNH₂ (10 mL) at 70 Cfor 2 h. The mixture was concentrated and diluted with conc. NH₄OH (5mL) and water (15 mL). Extraction (DCM, 80 mL) and concentration gaveN-methyl-4-(pent-4-ynyl)piperazine-1-carboxamide (0.659 g).

Step 4

Sonogashira coupling of4-chloro-5-iodo-7-((4-methoxy-3,5-dimethylpyridin-2-yl)methyl)-7H-pyrrolo[2,3-d]pyrimidin-2-amine withN-methyl-4-(pent-4-ynyl)piperazine-1-carboxamide according to thegeneral procedure A gave the title compound, as a solid. Mp=191.2-193.2°C. HPLC Rt=4.24 min.

Example 284-(5-(2-amino-4-chloro-7-((4-methoxy-3,5-dimethylpyridin-2-yl)methyl)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)pent-4-ynyl)-N-ethylpiperazine-1-carboxamide

Step 1: N-ethyl-4-(pent-4-ynyl)piperazine-1-carboxamide

A solution of 1-(pent-4-ynyl)piperazine (1.08) in DCM (5 mL) was treatedwith ethyl isocyanate (0.56 mL) at rt for 2 h, and evaporated to yieldN-ethyl-4-(pent-4-ynyl)piperazine-1-carboxamide.

Step 2

Sonogashira coupling of4-chloro-5-iodo-7-((4-methoxy-3,5-dimethylpyridin-2-yl)methyl)-7H-pyrrolo[2,3-d]pyrimidin-2-amine withN-ethyl-4-(pent-4-ynyl)piperazine-1-carboxamide according to the generalprocedure A gave the title compound, as a solid. Mp=200.5-203.3° C. HPLCRt=4.35 min.

Example 294-chloro-7-((4-methoxy-3,5-dimethylpyridin-2-yl)methyl)-5-(4-(4-phenylpiperazin-1-yl)but-1-ynyl)-7H-pyrrolo[2,3-d]pyrimidin-2-amine

Step 1: 1-(but-3-ynyl)-4-phenylpiperazine

A solution of but-3-ynyl 4-methylbenzenesulfonate (1.72 g), N-phenylpiperazine (1.17 mL) and diisoproylethylamine (1.45 mL) in1,2-dichloroethane (5 mL) was heated to reflux overnight. The mixturewas concentrated, diluted with sat. aq. NaHCO₃ (5 mL) and extracted withDCM (2×50 mL). Drying (Na₂SO₄) and silica gel flash chromatography (3%Et₃N in EtOAc) afforded 1-(but-3-ynyl)-4-phenylpiperazine (0.76 g)

Step 2

Sonogashira coupling of4-chloro-5-iodo-7-((4-methoxy-3,5-dimethylpyridin-2-yl)methyl)-7H-pyrrolo[2,3-d]pyrimidin-2-amine with 1-(but-3-ynyl)-4-phenylpiperazineaccording to the general procedure A gave the title compound, as asolid. HPLC Rt=4.97 min.

Example 304-chloro-7-((4-methoxy-3,5-dimethylpyridin-2-yl)methyl)-5-(4-(4-(pyridin-2-yl)piperazin-1-yl)but-1-ynyl)-7H-pyrrolo[2,3-d]pyrimidin-2-amine

Step 1: 1-(but-3-ynyl)-4-(pyridin-2-yl)piperazine

A solution of but-3-ynyl 4-methylbenzenesulfonate (1.27 g),1-(pyridin-2-yl)piperazine (1.17 g) and diisoproylethylamine (1.37 mL)in 1,2-dichloroethane (7 mL) was heated to reflux overnight. The mixturewas concentrated, diluted with sat. aq. NaHCO₃ (5 mL) and extracted withDCM (2×50 mL). Drying (Na₂SO₄) and silica gel flash chromatography(EtOAc/hexane 1:1→1:0) afforded1-(but-3-ynyl)-4-(pyridin-2-yl)piperazine (0.59 g)

Step 2

Sonogashira coupling of4-chloro-5-iodo-7-((4-methoxy-3,5-dimethylpyridin-2-yl)methyl)-7H-pyrrolo[2,3-d]pyrimidin-2-amine with 1-(but-3-ynyl)-4-(pyridin-2-yl)piperazineaccording to the general procedure A gave the title compound, as asolid. HPLC Rt=4.09 min.

Example 314-chloro-7-((4-methoxy-3,5-dimethylpyridin-2-yl)methyl)-5-(4-(4-(pyrimidin-2-yl)piperazin-1-yl)but-1-ynyl)-7H-pyrrolo[2,3-d]pyrimidin-2-amine

Step 1: 2-(4-(but-3-ynyl)piperazin-1-yl)pyrimidine

A solution of but-3-ynyl 4-methylbenzenesulfonate (1.14 g),2-(piperazin-1-yl)pyrimidine (1.06 g) and diisoproylethylamine (1.24 mL)in 1,2-dichloroethane (6 mL) was heated to reflux overnight. The mixturewas concentrated, diluted with sat. aq. NaHCO₃ (5 mL) and extracted withDCM (2×50 mL). Drying (Na₂SO₄) and silica gel flash chromatography(EtOAc/hexane 1:1→1:0) afforded2-(4-(but-3-ynyl)piperazin-1-yl)pyrimidine (0.60 g).

Step 2

Sonogashira coupling of4-chloro-5-iodo-7-((4-methoxy-3,5-dimethylpyridin-2-yl)methyl)-7H-pyrrolo[2,3-d]pyrimidin-2-amine with 2-(4-(but-3-ynyl)piperazin-1-yl)pyrimidineaccording to the general procedure A gave the title compound, as asolid. HPLC Rt=4.50 min.

Example 324-chloro-7-((4-methoxy-3,5-dimethylpyridin-2-yl)methyl)-5-(5-(4-phenylpiperazin-1-ylopent-1-ynyl)-7H-pyrrolo[2,3-d]pyrimidin-2-amine

Step 1: 1-(pent-4-ynyl)-4-phenylpiperazine

A solution of pent-4-ynyl methanesulfonate (1.05 g), N-phenyl piperazine(1.02 g) and diisoproylethylamine (1.2 mL) in THF (5 mL) was heated toreflux overnight. The mixture was concentrated, diluted with aq. NaOH 2M(10 mL) and extracted with DCM (2×50 mL). Drying (Na₂SO₄) and silica gelflash chromatography (EtOAc/hexane 2:1→2:0) afforded1-(pent-4-ynyl)-4-phenylpiperazine (1.03 g).

Step 2

Sonogashira coupling of4-chloro-5-iodo-7-((4-methoxy-3,5-dimethylpyridin-2-yl)methyl)-7H-pyrrolo[2,3-d]pyrimidin-2-amine with 1-(pent-4-ynyl)-4-phenylpiperazineaccording to the general procedure A gave the title compound, as asolid. HPLC Rt=5.08 min.

Example 334-chloro-7-((4-methoxy-3,5-dimethylpyridin-2-yl)methyl)-5-(5-(4-(pyridin-2-yl)piperazin-1-yl)pent-1-ynyl)-7H-pyrrolo[2,3-d]pyrmidin-2-amine

Step 1: 1-(pent-4-ynyl)-4-(pyridin-2-yl)piperazine

A solution of pent-4-ynyl methanesulfonate (1.05 g),1-(pyridin-2-yl)piperazine (1.02 g) and diisoproylethylamine (1.2 ml) inTHF (5 mL) was heated to reflux overnight. The mixture was concentrated,diluted with aq. NaOH 2M (10 mL) and extracted with DCM (2×50 mL).Drying (Na₂SO₄) and silica gel flash chromatography (EtOAc/hexane2:1→2:0) afforded 1-(pent-4-ynyl)-4-(pyridin-2-yl)piperazine (1.02 g).

Step 2

Sonogashira coupling of4-chloro-5-iodo-7-((4-methoxy-3,5-dimethylpyridin-2-yl)methyl)-7H-pyrrolo[2,3-d]pyrimidin-2-amine with 1-(pent-4-ynyl)-4-(pyridin-2-yl)piperazineaccording to the general procedure A gave the title compound, as asolid. HPLC Rt=4.23 min.

Example 344-chloro-7-((4-methoxy-3,5-dimethylpyridin-2-yl)methyl)-5-(5-(4-(pyrimidin-2-yl)piperazin-1-yl)pent-1-ynyl)-7H-pyrrolo[2,3-d]pyrimidin-2-amine

Step 1: 2-(4-(pent-4-ynyl)piperazin-1-yl)pyrimidine

A solution of pent-4-ynyl methanesulfonate (1.05 g),2-(piperazin-1-yl)pyrimidine (1.02 g) and diisoproylethylamine (1.2 mL)in THF (5 mL) was heated to reflux overnight. The mixture wasconcentrated, diluted with aq. NaOH 2M (10 mL) and extracted with DCM(2×50 mL). Drying (Na₂SO₄) and silica gel flash chromatography(EtOAc/hexane 2:1→2:0) afforded2-(4-(pent-4-ynyl)piperazin-1-yl)pyrimidine (0.95 g).

Step 2

Sonogashira coupling of4-chloro-5-iodo-7-((4-methoxy-3,5-dimethylpyridin-2-yl)methyl)-7H-pyrrolo[2,3-d]pyrimidin-2-amine with2-(4-(pent-4-ynyl)piperazin-1-yl)pyrimidine according to the generalprocedure A gave the title compound, as a solid. HPLC Rt=4.61 min.

Example 355-(5-(1H-imidazol-1-yl)pent-1-ynyl)-4-chloro-7-((4-methoxy-3,5-dimethylpyridin-2-yl)methyl)-7H-pyrrolo[2,3-d]pyrimidin-2-amine

Step 1: 1-(pent-4-ynyl)-1H-imidazole

A mixture of pent-4-ynyl methanesulfonate (1.06 g), imidazole (534 mg)and K₂CO₃ (3.66 g) in 2-butanone (11 mL) was heated to reflux overnight.Work-up (DCM, H₂O), drying (Na₂SO₄) and concentration gave1-(pent-4-ynyl)-1H-imidazole (472 mg).

Step 2

Sonogashira coupling of4-chloro-5-iodo-7-((4-methoxy-3,5-dimethylpyridin-2-yl)methyl)-7H-pyrrolo[2,3-d]pyrimidin-2-amine with 1-(pent-4-ynyl)-1H-imidazole according tothe general procedure A gave the title compound, as a solid. HPLCRt=4.40 min.

Example 364-(2-amino-4-chloro-7-(4-methoxy-3,5-dimethylpyridin-2-yl)methyl)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)but-3-ynylethyl carbonate

A solution of4-[2-Amino-4-chloro-7-(4-methoxy-3,5-dimethyl-pyridin-2-ylmethyl)-7H-pyrrolo[2,3-d]pyrimidin-5-yl]-but-3-yn-1-ol (see example 2) (1.53 g) and NaH(95%, 0.38 g) in anhydrous DMA (25 mL) was treated with ethylchloroformate (1.5 mL) at 0-23° C. for 0.5 h. Work-up and preparativeHPLC gave the title compound. HPLC Rt=5.65 min.

Example 37Ethyl-4-chloro-5-(4-(ethoxycarbonyloxy)but-1-ynyl)-7-((4-methoxy-3,5-dimethylpyridin-2-yl)methyl)-7H-pyrrolo[2,3-d]pyrimidin-2-ylcarbamate

A suspension of4-[2-Amino-4-chloro-7-(4-methoxy-3,5-dimethyl-pyridin-2-ylmethyl)-7H-pyrrolo[2,3-d]pyrimidin-5-yl]-but-3-yn-1-ol (see example 2) (0.73 g) inanhydrous pyridine (5 mL) and DCM (25 mL) was treated with ethylchloroformate (1 mL) at rt for 30 min. The mixture was cooled to 0° C.,treated with addition EtOCOCl (1 mL), and the temperature was allowed toslowly reach rt overnight. The reaction mixture was diluted with DCM (30mL) and washed sequentially with H₂O, NH₄OH 1M, and brine. The DCM wasevaporated, but not the residual pyridine, Addition of MeOH (20 mL)induced crystallization of the product, as white needles. HPLC Rt=6.28min.

Example 384-(4-(2-amino-4-chloro-7-((4-methoxy-3,5-dimethylpyridin-2-yl)methyl)-7H-pyrrolo-[2,3-d]pyrimidin-5-yl)but-3-ynyl)-N-methylpiperazine-1-carboxamide

Step 1: 1-(but-3-ynyl)piperazine

A solution of but-3-ynyl 4-methylbenzenesulfonate (2.0 mL) andpiperazine (2.0 g) in EtOH (6 mL) was heated to reflux for 30 min. Themixture was concentrated, diluted with NaOH 2 M (8 mL) and extractedwith Et₂O (50 mL). Evaporation of the organic layer gave a 2:1 mixtureof mono and bis-alkylated piperazine (450 mg) which was discarded. Theaqueous layer was further extracted with DCM (100 mL) to give of1-(but-3-ynyl)piperazine (640 mg).

Step 2: 4-(but-3-ynyl)-N-methylpiperazine-1-carboxamide

A solution of 1-(but-3-ynyl)piperazine (445 mg) in THF (4 mL) wastreated with 4-nitrophenyl carbonochloridate (649 mg) at rt for 5 min. Aprecipitate formed immediately, and the suspension was treated with Et₃N(0.45 mL) to ensure complete reaction. The suspension was diluted withH₂O (2 mL), MeOH (2 mL) and 40% aq. MeNH₂ (4.0 mL) and stirred at rt for3 days. Extraction with DCM (50 mL), washing (NaOH 2M), drying (Na₂SO₄)and concentration gave 4-(but-3-ynyl)-N-methylpiperazine-1-carboxamide(370 mg).

Step 3

Sonogashira coupling of4-chloro-5-iodo-7-((4-methoxy-3,5-dimethylpyridin-2-yl)methyl)-7H-pyrrolo[2,3-d]pyrimidin-2-amine with4-(but-3-ynyl)-N-methylpiperazine-1-carboxamide according to the generalprocedure A gave the title compound, as a solid. HPLC Rt=4.10 min

Example 395-(4-(1H-imidazol-1-yl)but-1-ynyl)-4-chloro-7-((4-methoxy-3,5-dimethylpyridin-2-ylmethyl)-7H-pyrrolo[2,3-d]pyrimidin-2-amine

Step 1: 1-(but-3-ynyl)-1H-imidazole

A mixture of but-3-ynyl 4-methylbenzenesulfonate (1.04 g), imidazole(573 mg) and K₂CO₃ (3.88 g) in 2-butanone (10 mL) was heated to refluxovernight. Work-up (DCM, H₂O), drying (Na₂SO₄) and concentration gave1-(but-3-ynyl)-1H-imidazole (501 mg).

Step 2

Sonogashira coupling of4-chloro-5-iodo-7-((4-methoxy-3,5-dimethylpyridin-2-yl)methyl)-7H-pyrrolo[2,3-d]pyrimidin-2-amine with 1-(but-3-ynyl)-1H-imidazole according tothe general procedure A gave the title compound., as a solid. HPLCRt=4.21 min.

Example 40(S)-4-(2-amino-4-chloro-7-((4-methoxy-3,5-dimethylpyridin-2-yl)methyl)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)but-3-ynyl2-aminopropanoate

A solution of4-[2-Amino-4-chloro-7-(4-methoxy-3,5-dimethyl-pyridin-2-ylmethyl)-7H-pyrrolo[2,3-d]pyrimidin-5-yl]-but-3-yn-1-ol (see example 2) (51.7 mg), N-Bocalanine (53.3 mg), DMAP (33.7 mg), and EDCI (54.9 mg) in anhyrous DMA (4mL) was stirred at rt overnight. Work-up (EtOAc; H₂O) gave the crudeBoc-protected intermediate. A solution of this intermediate (40 mg) inDCM (1.0 (mL) was treated with TFA (0.2 mL) at rt for 5 min, andevaporated. The material was dissolved in MeOH (1 ml) and water (10 mL),and washed with Et₂O (10 mL). The organic layer was discarded. Theaqueous layer was brought to pH>7 with sat. aq. NaHCO₃ andback-extracted with DCM. The title compound was isolates as a solid.HPLC Rt=4.37 min.

Example 41(R)-4-(2-amino-4-chloro-7-((4-methoxy-3,5-dimethylpyridin-2-yl)methyl)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)but-3-ynyl2-amino-3-methylbutanoate

This compound was obtained by the same procedure as used for example 40,using N-Boc valine in place of N-Boc alanine, to give the title compoundas a solid. HPLC Rt=4.58 min.

Example 42(R)-4-(2-amino-4-chloro-7-((4-methoxy-3,5-dimethylpyridin-2-yl)methyl)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)but-3-ynyl2-amino-4-methylpenitanoate

This compound was obtained by the same procedure as used for example 40,using N-Boc isoleucine in place of N-Boc valine, to give the titlecompound as a solid. HPLC Rt=4.76 min.

Example 434-(2-amino-4-chloro-7-((4-methoxy-3,5-dimethylpyridin-2-yl)methyl)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)but-3-ynyldi-tert-butyl phosphate

A solution of4-[2-Amino-4-chloro-7-(4-methoxy-3,5-dimethyl-pyridin-2-ylmethyl)-7H-pyrrolo[2,3-d]pyrimidin-5-yl]-but-3-yn-1-ol (see example 2) (4.0 g) and1-H-tetrazole (0.45 M in acetonitrile, 70 mL) in THF (160 mL) wastreated with di-tert-butyl diisopropylphosphoramidite (16.4 mL) at rtfor 2 h. The solution was cooled to 0° C. and treated with 30% aqueousH₂O₂ (11 mL) for 30 min. Work-up (EtOAc, aq. NaHCO₃) and reverse-phasepreparative HPLC gave the title compound, as a solid. HPLC Rt=6.13 min.

Example 44 4-(2-amino-4-chloro-7-((4-methoxy-3,5-dimethylpyridin-2-yl)methyl)-7H-pyrrolo[2,3d]pyrimidin-5-yl)but-3-ynyl dihydrogen phosphate

A solution of4-(2-amino-4-chloro-7-((4-methoxy-3,5-dimethylpyridin-2-yl)methyl)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)but-3-ynyldi-tert-butyl phosphate (see example 43) in DCM was treated with TFA atrt for 1h. Evaporation gave the title compound as a TFA salt (5 g). Thecrude was dissolved in a mixture of Et₃N (0.7 mL and MeOH (60 mL),loaded on Dowex 50Wx2-400 (20 g, pre-washed with MeOH), and the resinwas washed with MeOH (200 mL) to remove the excess TFA. The desiredphosphate was released from the resin with Et₃N:MeOH 1:10 (200 mL) Thesolution was concentrated to give the phosphate as an oily triethylaminesalt (2.75 g). Crystallization was induced with EtOH (60 mL). Themixture was left at rt for 1.5 h and at −20° C. overnight to give afirst crop of crystals (0.64 g). The mixture was concentrated and thecrystallization procedure was repeated to give a second crop (0.74 g).The combined triethylamine salts (1274 mg) were dissolved in MeOH (50mL) with the help of Et₃N (326 μL) and treated with a 1.0 M solution ofNaOH in MeOH (4.68 mL). Evaporation and drying on high vacuum overnightgave the title compound as a solid sodium salt. HPLC Rt=4.21 min.

Example 452-((2-amino-4-chloro-5-(4-hydroxybut-1-ynyl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)methyl)-4-methoxy-3,5-dimethylpyridine1-oxide

Step 1:2-((4-chloro-5-iodo-2-pivalamido-7H-pyrrolo[2,3-d]pyrimidin-7-yl)methyl)-4-methoxy-3,5-dimethylpyridine 1-oxide

A mixture ofN-(4-chloro-5-iodo-7H-pyrrolo[2,3-d]pyrimidin-2-yl)pivalainide (235 mg),2-(chloromethyl)-4-methoxy-3,5-dimethylpyridine 1-oxide (129 mg), andK₂CO₃ (413 mg) in DMF (6.0 mL) was stirred at rt for 3 days, and dilutedwith water. The precipitate was collected by filtration and washed withIPA to give2-((4-chloro-5-iodo-2-pivalamido-7H-pyrrolo[2,3-d]pyrimidin-7-yl)methyl)-4-methoxy-3,5-dimethylpyridine1-oxide (124 mg). Solid. HPLC Rt=6.73 min

Step 2:2-((4-chloro-5-(4-hydroxybut-1-ynyl)-2-pivalamido-7H-pyrrolo[2,3-d]pyrimidin-7-yl)methyl)-4-methoxy-3,5-dimethylpyridine1-oxide

Sonogashira coupling of2-((4-chloro-5-iodo-2-pivalamido-7H-pyrrolo[2,3-d]pyrimidin-7-yl)methyl)-4-methoxy-3,5-dimethylpyridine1-oxide (335 mg) with but-3-yn-1-ol (200 μL) according to the generalprocedure A gave crude2-((4-chloro-5-(4-hydroxybut-1-ynyl)-2-pivalamido-7H-pyrrolo[2,3-d]pyrimidin-7-yl)methyl)-4-methoxy-3,5-dimethylpyridine1-oxide (291 mg).

Solid. HPLC Rt=5.82 min.

Step 3

Crude2-((4-chloro-5-(4-hydroxybut-1-ynyl)-2-pivalamido-7H-pyrrolo[2,3-d]pyrimidin-7-yl)methyl)-4-methoxy-3,5-dimethylpyridine1-oxide (291 mg) was dissolved in 10 mL EtOH. An aliquot of thissolution (2 mL) was further diluted with EtOH (5 mL) and treated withZnCl₂ (103 mg) at 120° C. for 20 min in a microwave oven. Reverse-phasepreparative HPLC gave the title compound (2.2 mg), as a solid. HPLCRt=4.88 min.

Example 465-(but-3-en-1-ynyl)-4-chloro-7-((4-methoxy-3,5-dimethylpyridin-2-yl)methyl)-7H-pyrrolo[2,3-d]pyrimidin-2-amine

A mixture of4-(2-amino-4-chloro-7-((4-methoxy-3,5-dimethylpyridin-2-yl)methyl)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)but-3-ynyl 4-methylbenzenesulfonate (516 mg) andCsF (2.0 g) in DMF (7.0 mL) was heated to 70° C. for 1 h. Work-up andflash chromatography (1→3% MeOH in DCM) gave the title compound (280mg), as a pale yellow oil. HPLC Rt=5.90 min.

Example 475-(2-amino-4-chloro-7-((4-methoxy-3,5-dimethylpyridin-2-yl)methyl)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)pent-4-ynylmethanesulfonate

Sonogashira coupling of4-chloro-5-iodo-7-((4-methoxy-3,5-dimethylpyridin-2-yl)methyl)-7H-pyrrolo[2,3-d]pyrimidin-2-amine with pent-4-ynyl methanesulfonate according tothe general procedure A gave the title compound, as a solid. HPLCRt=5.31 min.

Example 484-chloro-5-(5-fluoropent-1-ynyl)-7-((4-methoxy-3,5-dimethylpyridin-2-yl)methyl)-7H-pyrrolo[2,3-d]pyrimidin-2-amine

A solution of KF (77.8 mg), Kryptofix 222 (Aldrich, 839 mg) and water(10 mL) was evaporated to dryness, and dried first azeotropically withanh. CH₃CN (3×20 mL), then on high vacuum at 50° C. overnight. The solidwas redissolved in anhydrous acetonitrile (13 mL) to give a 0.1 Mfluoride solution. Treatment of5-(2-amino-4-chloro-7-((4-methoxy-3,5-dimethylpyridin-2-yl)methyl)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)pent-4-ynyl methanesulfonate (see example 47) (42mg) with this solution (3 mL) at 60-70° C. for 2 h, work-up andreverse-phase preparative HPLC gave the title compound, as an oil. HPLCRt=5.88 min.

Example 49(S)-4-(2-amino-4-chloro-7-((4-methoxy-3,5-dimethylpyridin-2-yl)methyl)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)but-3-yne-1,2-diol

Treatment of5-(but-3-en-1-ynyl)-4-chloro-7-((4-methoxy-3,5-dimethylpyridin-2-yl)methyl)-7H-pyrrolo[2,3-d]pyrimidin-2-amine (see example 46) (35 mg) with AD-mix-α(Aldrich, 540 mg) in t-BuOH:THF:water 1:1:1 (6 mL) at rt overnight,followed by work-up and reverse-phase preparative HPLC gave the titlecompound (10 mg), as a solid. HPLC Rt=4.13 min

Example 50(R)-4-(2-amino-4-chloro-7-((4-methoxy-3,5-dimethylpyridin-2-yl)methyl)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)but-3-yne-1,2-diol

Treatment of5-(but-3-en-1-ynyl)-4-chloro-7-((4-methoxy-3,5-dimethylpyridin-2-yl)methyl)-7H-pyrrolo[2,3-d]pyrimidin-2-amine (see example 46) (35 mg) with AD-mix-β(Aldrich, 540 mg) in t-BuOH:THF:water 1:1:1 (6 mL) at rt overnight,followed by work-up and reverse-phase preparative HPLC gave the titlecompound (10 mg), as a solid. HPLC Rt=4.13 min

Example 514-chloro-7-((4-methox-3,5-dimethylpyridin-2-yl)methyl)-5-(3-methylbut-3-en-1-ynyl)-7H-pyrrolo[2,3-d]pyrimidin-2-amine

Sonogashira coupling of4-chloro-5-iodo-7-((4-methoxy-3,5-dimethylpyridin-2-yl)methyl)-7H-pyrrolo[2,3-d]pyrimidin-2-amine with 2-methylbut-1-en-3-yne according to thegeneral procedure

A gave the title compound, as a solid. HPLC Rt=6.00 min.

Example 52(S)-4-(2-amino-4-chloro-7-((4-methoxy-3,5-dimethylpyridin-2-yl)methyl)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-methylbut-3-yne-1,2-diol

Treatment of4-chloro-7-(1-methoxy-3,5-dimethylpyridin-2-yl)methyl)-5-(3-methylbut-3-en-1-ynyl)-7H-pyrrolo[2,3-d]pyrimidin-2-amine(see example 51) (263 mg) with AD-mix-α (Aldrich, 1.41 g) int-BuOH:water 1:1: (7 mL) at rt for 3 days, followed by quench (NaHSO₃,1.6 g), work-up, and preparative HPLC gave the title compound (69 mg),as a solid. HPLC Rt=4.36 min

Example 53(R)-4-(2-amino-4-chloro-7-((4-methoxy-3,5-dimethylpyridin-2-yl)methyl)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-methylbut-3-yne-1,2-diol

Treatment of4-cloro-7-((4-methoxy-3,5-dimethylpyridin-2-yl)methyl)-5-(3-methylbut-3-en-1-ynyl)-7H-pyrrolo[2,3-d]pyrimidin-2-amine(see example 51) (357 mg) with AD-mix-β (Aldrich, 1.57 g) int-BuOH:water 1:1: (10 mL) at rt for 18 h, followed by quench (NaHSO₃,1.2 g), work-up, and silica gel flash chromatography (1→10% MeOH in DCM)gave the title compound (61 mg) and recovered starting material (263mg). Solid. HPLC Rt=4.36 min.

Example 54(R)—((S)-4-(2-amino-4-chloro-7-((4-methoxy-3,5-dimethylpyridin-2-yl)methyl)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-hydroxy-2-methylbut-3-ynyl)3,3,3-trifluoro-2-methoxy-2-phenylpropanoate

A mixture of(S)-4-(2-amino-4-chloro-7-((4-methoxy-3,5-dimethylpyridin-2-yl)methyl)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-methylbut-3-yne-1,2-diol (see example 52) (18mg), (S)-(+)-α-methoxy-alpha-(trifluoromethyl)phenylacetyl chloride (45μL), and Et₃N (50 μL) in THF (3.0 mL) was heated to 70° C. for 2 h, andthen to 40° C. overnight. Preparative TLC (EtOAc/DCM 25:65) gave thetitle compound, as a solid. HPLC Rt=6.45 min.

Example 55(R)—((R)-4-(2-amino-4-chloro-7-((4-methoxy-3,5-dimethylpyridin-2-yl)methyl)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-hydroxy-2-methylbut-3-ynyl)3,3,3-trifluoro-2-methoxy-2-phenylpropanoate

A mixture of(R)-4-(2-amino-4-chloro-7-((4-methoxy-3,5-dimethylpyridin-2-yl)methyl)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-methylbut-3-yne-1,2′-diol (see example 53) (22mg), (S)-(+)-α-methoxy-alpha-(trifluoromethyl)phenylacetyl chloride (45μL), and Et₃N (50 μL) in THF (3.0 mL) was heated to 40° C. overnight.Preparative TLC (EtOAc/DCM 25:65) gave the title compound, as a solid.HPLC Rt=6.44 min.

Example 562-((2-amino-4-chloro-7-((4-methoxy-3,5-dimethylpyridin-2-yl)methyl)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)ethynyl)-2-hydroxypropane-1,3-diyldiacetate

Step 1: 2-ethynyl-2-hydroxypropane-1,3-diyl diacetate

An ethynylmagnesium bromide solution (0.5 M in THF) was cooled to −78°C. under nitrogen. A solution of 1,3-diacetoxyacetone (0.5 g) in THF(3.0 mL) was added, and the resulting mixture was stirred for 30 min at−78° C., then at 0° C. for 5 min, quenched with 1.0 N aq. HCl (3 mL) andextracted in EtOAc. Evaporation gave crude2-ethynyl-2-hydroxypropane-1,3-diyl diacetate, contaminated with about50 mol % of 1,3-dacetoxyacetone. The crude material was used withoutfurther purification.

Step 2

Sonogashira coupling of4-chloro-5-iodo-7-((4-methoxy-3,5-dimethylpyridin-2-yl)methyl)-7H-pyrrolo[2,3-d]pyrimidin-2-amine with crude 2-ethynyl-2-hydroxypropane-1,3-diyldiacetate according to the general procedure A gave the title compound,as an oil. HPLC Rt=4.89 min.

Example 572-((2-amino-4-chloro-7-((4-methoxy-3,5-dimethylpyridin-2-yl)methyl)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)ethynyl)propane-1,2,3-triol

A solution of2-((2-amino-4-chloro-7-((4-methoxy-3,5-dimethylpyridin-2-yl)methyl)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)ethynyl)-2-hydroxypropane-1,3-diyl diacetate (seeexample 56) (60 mg) and Et₃N (1 mL) in THF (1 mL) and MeOH (6 ml,) washeated in a microwave oven to 150° C. for 20 min. Concentration andpreparative HPLC gave the title compound, as a solid. HPLC Rt =3.90 min.

Example 584-(2-amino-4-chloro-7-((4-methoxy-3,5-dimethylpyridin-2-yl)methyl)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-methylbut-3-yn-2-ol

Sonogashira coupling of4-chloro-5-iodo-7-((4-methoxy-3,5-dimethylpyrimidin-2-yl)methyl)-7H-pyrrolo[2,3-d]pyrimidin-2-amine with 2-methylbut-3-yn-2-ol according to thegeneral procedure A gave the title compound, as a solid. HPLC Rt=4.81min.

Example 595-(2-amino-4-chloro-7-((4-methoxy-3,5-dimethylpyridin-2-yl)methyl)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)pent-4-yn-2-ol

Sonogashira coupling of4-chloro-5-iodo-7-((4-methoxy-3,5-dimethylpyridin-2-yl)methyl)-7H-pyrrolo[2,3-d]pyrimidin-2-amine with pent-4-yn-2-ol according to the generalprocedure A gave the title compound, as a solid. HPLC Rt=4.29 min

Example 604-(2-amino-4-chloro-7-((4-methoxy-3,5-dimethylpyridin-2-yl)methyl)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)but-3-ynylacetate

Step 1: But-3-ynyl acetate

A solution of but-3-yn-1-ol (3.0 mL) and Et₃N (6.6 mL) in1,2-dichloroethane (10 mL) was treated with AcCl (3.1 mL) with caution(very exothermic). Work-up gave but-3-ynyl acetate.

Step 2

Sonogashira coupling of4-chloro-5-iodo-7-((4-methoxy-3,5-dimethylpyridin-2-yl)methyl)-7H-pyrrolo[2,3-d]pyrimidin-2-armine with but-3-ynyl acetate according to thegeneral procedure A gave the title compound, as a solid. HPLC Rt=5.31min.

Example 614-(2-amino-4-chloro-7-((4-methoxy-3,5-dimethylpyridin-2-yl)methyl)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)but-3-ynylsulfamate

A mixture of formic acid (0.463 g) and N,N-dimethylacetamide (1 drop)was added to a solution of chlorosulfonyl isocyanate (1.31 g) in DCM (9mL), and heated to reflux for 3 h. The resulting solution was added to asolution of4-[2-Amino-4-chloro-7-(4-methoxy-3,5-dimethyl-pyridin-2-ylmethyl)-7H-pyrrolo[2,3-d]pyrimidin-5-yl]-but-3-yn-1-ol(see example 2) (200 mg) in DMA (10 mL) and stirred at rt for 30 min.Work-up and reverse-phase preparative HPLC gave the title compound, as asolid. HPLC Rt=4.96 min.

Example 625-(2-amino-4-chloro-7-((4-methoxy-3,5-dimethylpyridin-2-yl)methyl)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)pent-4-ynylsulfamate

The title compound was obtained by the same procedure as in example 61,carrying out the reaction on5-[2-Amino-4-chloro-7-(4-methoxy-3,5-dimethyl-pyridin-2-ylmethyl)-7H-pyrrolo[2,3-d]pyrimidin-5-yl]-pent-4-yn-1-ol(see example 2) Solid. HPLC Rt=4.77 min.

Example 634-(2-amino-4-chloro-7-((4-methoxy-3,5-dimethylpyridin-2-yl)methyl)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)but-3-ynylhydrogen sulfate

Step 1: But-3-ynyl hydrogen sulfate, pyridine salt

But-3-yn-1-ol (2.1 g) in DCM (20 mL) was treated with sulfurtrioxide-pyridine complex (4.77 g) overnight. The solid was filtered offand the reaction mixture was concentrated and used without furtherpurification.

Step 2

Sonogashira coupling of4-chloro-5-iodo-7-((4-methoxy-3,5-dimethylpyridin-2-yl)methyl)-7H-pyrrolo[2,3-d]pyrimidin-2-amine with the pyridine salt of but-3-ynyl hydrogensulfate according to the general procedure A gave the title compound.

Solid. HPLC Rt=4.24 min.

Example 644-(2-amino-4-chloro-7-((4-methoxy-3,5-dimethylpyridin-2-yl)methyl)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)but-3-ynylbis(2,2,2-trichloroethyl)phosphate

Step 1: But-3-ynyl bis(2,2,2-trichloroethyl) phosphate

A solution of but-3-yn-1-ol (0.413 g) in DCE (20 mL) was treated withEt₃N (2 mL) and bis(2,2,2-trichloroethyl)phosphorochloride (2.27 g) at50° C. overnight. Work-up gave crude but-3-ynylbis(2,2,2-trichloroethyl) phosphate as an oil that was used withoutfurther purification.

Step 2

Sonogashira coupling of4-chloro-5-iodo-7-((4-methoxy-3,5-dimethylpyridin-2-yl)methyl)-7H-pyrrolo[2,3-d]pyrimidin-2-amine with but-3-ynyl bis(2,2,2-trichloroethyl)phosphate according to the general procedure I gave the title compound.

Solid. HPLC Rt=6.69 min.

Example 65 rHSP90 Competitive Binding Assay

Five microgram of purified rHSP90 protein (Stressgen, BC, Canada,#SPP-770) in phosphate buffered saline (PBS) was coated on 96 wellplates by incubating overnight at 4° C. Unbound protein was removed andthe coated wells were washed twice with 200 μL PBS. DMSO controls(considered as untreated samples) or test compounds were then added at100-30-10-3-1-0.3 μM dilutions (in PBS), the plates mixed for 30 secondson the plate shaker, and then incubated for 60-min. at 37° C. The wellswere washed twice with 200 μL PBS, and 10 μM biotinylated-geldanamycin(biotin-GM) was added and incubated for 60 min. at 37° C. The wells werewashed again twice with 200 μL PBS, before the addition of 20 μg/mLstreptavidin-phycoerythrin (streptavidin-PE) (Molecular Probes, Eugene,Oreg.) and incubation for 60 min. at 37° C. The wells were washed againtwice with 200 μL PBS. Relative fluorescence units (RFU) was measuredusing a SpectraMax Gemini XS Spectrofluorometer (Molecular Devices,Sunnyvale, Calif.) with an excitation at 485 nm and emission at 580 nm;data was acquired using SOFTmax® PRO software (Molecular DevicesCorporation, Sunnyvale, Calif.). The background was defined as the RFUgenerated from wells that were not coated with HSP90 but were treatedwith the biotin-GM and streptavidin-PE. The background measurements weresubtracted from each sample treated with biotin-GM and streptavidin-PEmeasurements before other computation. Percent inhibition of binding foreach sample was calculated from the background subtracted values asfollows:% binding inhibition=[(RFU untreated−RFU treated)/RFU untreated]×100.

Example 66

Cell Lysate Binding Assay

MCF7 breast carcinoma cell lysates were prepared by douncing in lysingbuffer (20 mM HEPES, pH 7.3, 1 mM EDTA, 5 mM MgCl₂, 100 mM KCl), andthen incubated with or without test compound for 30 mins at 4° C.,followed by incubation with biotin-GM linked to BioMag™ streptavidinmagnetic beads (Qiagen) for 1 hr at 4° C. The tubes were placed on amagnetic rack, and the unbound supernatant removed. The magnetic beadswere washed three times in lysis buffer and boiled for 5 mins at 95° C.in SDS-PAGE sample buffer. Samples were analyzed on SDS protein gels,and Western Blots were done for rHSP90. Bands in the Western Blots werequantitated using the Bio-rad Fluor-S MultiImager, and the % inhibitionof binding of rHSP90 to the biotin-GM was calculated. The lysate bindingability of selected compounds of the invention based on the above assayis summarized in Table 3. The IC₅₀ reported is the concentration of testcompound needed to achieve 50% inhibition of the biotin-GM binding torHSP90 in the MCF7 cell lysates.

Example 67 Her2 Degradation Assay

Many cancers are associated with the over expression Her2 protein. Ithas been shown that compounds able to diminish Her2 levels show goodpromise as anti-cancer agents. Thus a good in vitro assay foridentifying compounds of the present invention, which are likely todemonstrate anti-cancer activity, is the Her2 degradation assay, asdescribed below:

MCF7 breast carcinoma cells (ATCC) were grown in Dulbecco's modifiedEagle's medium (DMEM) containing 10% fetal bovine serum (FBS) and 10 mMHEPES, and plated in 24 well plates (50% confluent). Twenty-four hrslater (cells are 65-70% confluent), test compounds were added andincubated overnight for 16 h. For the less potent compounds, the amountsadded were 100 μM, 30 μM, 10 μM and 1 μM, and for more potent compounds,the amounts added were 1 μM, 0.3 μM, 0.1 μM, 0.03 μM, 0.01 μM and 0.003M. The wells were washed with 1 mL phosphate buffered saline (PBS), and200 μL trypsin was added to each well. After trypsinization wascomplete, 50 μL of FBS was added to each well. Then 200 μL cells weretransferred to 96 well plates. The cells were pipetted up and down toobtain a single cell suspension. The plates were centrifuged at 2,500rpm for 1 min using a Sorvall Legend RT™ tabletop centrifuge (KendroLaboratory Products, Asheville, N.C.). The cells were then washed oncein PBS containing 0.2% BSA and 0.2% sodium azide (BA buffer).Phycoerythrin (PE) conjugated anti IER2/Neu antibody (Becton Dickinson,#340552), or PE conjugated anti-keyhole limpet hemocyanin [KLH] (BectonDickinson, #340761) control antibody was added at a dilution of 1:20 and1:40 respectively (final concentration was 1 μg/mL) and the cells werepipeted up and down to form a single cell suspension, and incubated for15 mins. The cells were washed twice with 200 μL BA buffer, andresuspended in 200 μL BA buffer, and transferred to FACSCAN tubes withan additional 250 μL BA buffer. Samples were analyzed using aFACSCalibur™ flow cytometer (Becton Dickinson, San Jose, Calif.)equipped with Argon-ion laser that emits 15 mW of 488 nm light forexcitation of the PE fluorochrome. 10,000 events were collected persample. A fluorescence histogram was generated and the mean fluorescenceintensity (MFI) of each sample was determined using Cellquest software.The background was defined as the NM generated from cells incubated withcontrol IgG-PE, and was subtracted from each sample stained with theHER2/Neu antibody. Cells incubated with DMSO were used as untreatedcontrols since the compounds were resuspended in DMSO. Percentdegradation of Her2 was calculated as follows:% Her2 degraded=[(MFI untreated cells−MFI treated cells)/MFI untreatedcell]×100.

The Her2 degradation ability of selected compounds of the inventionbased on this assay is summarized in Table 3. IC₅₀ is defined as theconcentration at which there was 50% degradation of the HER2/Neuprotein.

Example 68 MTS Assay

3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazoliumassays measure the cytotoxicity of geldanamycin derivatives. MTS(3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium) is atetrazolium dye that is converted to a formazan product by dehydrogenaseenzymes of metabolically active cells (Corey, A. et al. “Use of anaqueous soluble tetrazolium/formazan assay for cell growth assays inculture,” Cancer Commun. 1991, 3, 207-212). Cells were seeded in 96 wellplates at 2000 cells/well and allowed to adhere overnight in Dulbecco'smodified Eagle's medium supplemented with 10% fetal bovine serum. Thefinal culture volume was 100 μl. Viable cell number was determined byusing the Celltiter 96 AQ_(ueous) Non-radioactive Cell ProliferationAssay (Promega, Madison Wis.). The MTS/PMS (phenazine methosulfate)solution was mixed at a ratio of 20:1, and 20 μL was added per well to100 μl of culture medium. After 2-4 hours, the formation of the formazanproduct was measured at 490 nm absorbance using a multiwell platespectrophotometer. Background was determined by measuring the Abs 490 nmof cell culture medium and MTS-PMS in the absence of cells and wassubtracted from all values. Percent viable cells was calculated asfollows:% viable cells=(Abs at 490 nm treated cells/Abs at 490 nm untreatedcells)×100

The effect of selected compounds of the invention on MCF7 breastcarcinoma cells according to the MTS assay is summarized in table 3.IC₅₀ was defined as the concentration of the compound which gave rise to50% reduction in viable cell number.

TABLE 3 Biological Activities of Selected Compounds of the Invention

IC₅₀ (μM) Sample Lysate MTS MTS No Example R³ Her2 binding BT474 MCF7 11 —CH₂OH 0.012 ND 0.02 0.13 2 10 —CH₂CH₂CON(Et)₂ 0.03 0.018 0.05 0.7 3 2—CH₂CH₂OH 0.006 0.003 0.006 0.013 4 3 —CH₂CH₂CH₂OH 0.006 0.003 0.0080.013 5 5 —CH₂N(iPr)₂ 0.055 ND 0.08 0.1 6 11

0.018 ND 0.02 0.1 7 12 —CH₂CH₂CONH₂ 0.016 0.012 0.02 0.03 8 16—CH₂CH₂CH₂NH₂ 0.028 ND >10 0.09 9 15 —CH₂CH₂CH₂NHBOC 0.009 ND 1.0 0.0610 17

0.007 ND 0.1 0.01 11 18

0.01 ND 1.0 0.05 12 19

0.015 ND 1.0 0.1 13 20 —CH₂CH₂NHtBu 0.012 ND 23 130 14 21—CH₂CH₂CH₂NHt-Bu 0.015 ND 30 230 15 7 —CH₂CH₂CH₂CH₂OH 0.009 ND 20 200 1622 —CH₂O(CO)CH₂NMe₂ 0.013 ND 20 200 ND, not determined.

Example 69 In Vivo Mouse Tumor Studies

Six to 8 week old Balb/C and nu/nu athymic female mice were obtainedfrom Harlan Sprague Dawley, (Indianapolis, Ind.). The mice weremaintained in sterilized filter topped cages or ventilated caging in aroom with a 12 hour light/dark cycle. Irradiated pelleted food (HarlanTeklad #7912) and autoclaved deionized water were provided ad libitum.Animals were identified by the use of individually numbered ear tags.Experiments were carried out under institutional guidelines for theproper and human use of animals in research established by the Institutefor Laboratory Animal Research (ILAR).

Tumor fragments (approximately 2 mm³) or 5×10⁶ tumor cells wereinoculated subcutaneously in the right or left flank of the animal. Micewith established tumors (50-200 mm³) were selected for study(n=7-10/treatment group). Tumor dimensions were measured using calipersand tumor volumes were calculated using the equation for an ellipsoidsphere (1×w²)/2=mm³, where 1 and w refer to the larger and smallerdimensions collected at each measurement.

Mice were followed until tumor volumes in the control group reachedapproximately 1000 mm³ and were sacrificed by CO₂ euthanasia. The meantumor volumes of each group were calculated. The change in mean treatedtumor volume was divided by the change in mean control tumor volume,multiplied by 100 and subtracted from 100% to give the tumor growthinhibition for each group. Statistical analysis was performed using thestandard T-test and using GraphPad Prism© Software.

Example 70 N87 Gastric Carcinoma Xenograft

Tumors were established in mice, by innoculation of human N87 stomachcancer cells, according to example 69. Four treatment groups(n=7-10/treatment group) were established for study:

-   I.    4-[2-Amino-4-chloro-7-(4-methoxy-3,5-dimethyl-pyridin-2-ylmethyl)-7H-pyrrolo[2,3-d]pyrimidin-5-yl]-but-3-yn-1-ol,    prepared as described in example 2 above, was administered at 8    mg/kg po-   II.    5-(2-amino-4-chloro-7-((4-methoxy-3,5-dimethylpyridin-2-yl)methyl)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-methylpent-4-yn-2-ol,    prepared as described in example 23 above, was administered at 8    mg/kg po-   III.    5-(2-amino-4-chloro-7-((4-methoxy-3,5-dimethylpyridin-2-yl)methyl)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-methylpent-4-yn-2-ol,    prepared as described in example 23 above, was administered at 16    mg/kg po-   IV. Vehicle alone was administered to the control group.

The results are shown in FIG. 1, a plot of tumor volume (mm³) againsttime (days).

Example 71 NC1295 Adrenocortical Carcinoma Xenograft

Tumors were established in mice, by innoculation of NCI295adrenocortical carcinoma cells, according to example 69. Two treatmentgroups (n=7-10/treatment group) were established for study:

-   I.    5-(2-amino-4-chloro-7-((4-methoxy-3,5-dimethylpyridin-2-yl)methyl)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-methylpent-4-yn-2-ol,    prepared as described in example 23 above, was administered at 8    mg/kg po-   II. Vehicle alone was administered to the control group.

The results are shown in FIG. 2, a plot of tumor volume (mm³) againsttime (days).

Example 72 SK-MEL-28 Melanoma Xenograft

Tumors were established in mice, by innoculation of ISK-MEL-28 melanomacells, according to example 69. Two treatment groups (n=7-10/treatmentgroup) were established for study:

-   I.    5-(2-amino-4-chloro-7-((4-methoxy-3,5-dimethylpyridin-2-yl)methyl)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-methylpent-4-yn-2-ol,    prepared as described in example 23 above, was administered at 8    mg/kg po-   II. Vehicle alone was administered to the control group.

The results are shown in FIG. 3, a plot of tumor volume (mm³) againsttime (days).

Example 73 HT29 Colon Carcinoma Xenograft

Tumors were established in mice, by innoculation of HIT29 ColonCarcinoma cells, according to example 69. Three treatment groups(n=7-10/treatment group) were established for study:

-   I.    5-(2-amino-4-chloro-7-((4-methoxy-3,5-dimethylpyridin-2-yl)methyl)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-methylpent-4-yn-2-ol,    prepared as described in example 23 above, was administered at 4    mg/kg po-   II.    5-(2-amino-4-chloro-7-((4-methoxy-3,5-dimethylpyridin-2-yl)methyl)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-methylpent-4-yn-2-ol,    prepared as described in example 23 above, was administered at 8    mg/kg po-   III. Vehicle alone was administered to the control group.

The results are shown in FIG. 4, a plot of tumor volume (mm³) againsttime (days).

Example 74 Pharmacodynamic Effects

Preparation of Buffers and Reagents

The following reagents and buffers are prepared ahead of theexperiments.

Western Lysis Buffer (WLS) contains 10 mM HEPES, 42 mM KCl, 5 mM MgCl2,0.1mM EDTA, 0.1 mM EGTA, 1 mM DTT, 1 mM PMSF, 1 ug/ml Pepstatin A, 1ug/ml Leupeptin, 5 ug/ml Aprotinin, and 1% Triton X-100. Aliquot andstore at −20° C.

5× Western Sample Buffer (5×WSB) contains 20 ml glycerol, 4 mlβ-mercaptoethanol, 5 g SDS, 12.5 ml 1M Tris pH 6.8, and 50 mgbormophenol blue. Add water to a final volume of 50 ml. Aliquot andstore at −20° C.

Western Transfer Buffer contains 23.3 g Tris base, 116 g glycine and 1.6L methanol. Add water to a final volume of 8 L and store at 4° C.

TBST (1×) contains 10 mM Tris pH8.0, 150 mM NaCl and 0.1% Tweene 20.

Blocking solution contains 5% nonfat dry milk in 1×TBST. Keep at 4° C.

Processing of Tumor Samples:

Snap-frozen N87 gastric carcinoma tumors are transferred from liquidnitrogen into −80° C. freezer. WLB is supplemented with proteaseinhibitor cocktail (stock at 100×) on ice. Each tumor is thawed on iceand transferred onto the lid of a Petri dish. It is covered with 50 ulof WLB and is dissected into smaller pieces with disposable scalpels.Any residual skin attached to the tumor is removed. The tumor pieces arethen chopped further down, and transferred into 300-500 ul of ice-coldVLB. The minced sample is sonicated at setting 3 on Fisher Scientific'sSonic Dismembrator 550 until no more solid pieces can be broken down.The suspension is then centrifuged at 15,000 g, 4° C. for 5 minutes. Thesupernatant is collected into a clean Eppendorf tube on ice as lysate. 2ul of the lysate is used for total protein quantification by followingdirections in the BCA

Protein Assay kit. The rest of the lysate is snap-frozen in liquidnitrogen while the BCA assay is in process. The total proteinconcentration in each lysate is calculated upon completion of BCA assay.The lysates are thawed in a water bath, and their total proteinconcentrations adjusted to 4-10 mg/ml using 5×WSB (to a final of 20% ofthe total volume) and WLB (if necessary). The adjusted lysates areboiled at 95° C. for 5 minutes and cooled to room temperature. At thisstage, they are frozen at −20° C. for future Western blotting analysis.

Western Blotting Analysis:

The quantified and adjusted lysates from tumor or spleen samples arethawed in a water bath, and loaded at equal total protein amount onto4-12% Tris-glycine precast gels together with biotinylated proteinmarker. Electrophoresis is carried out at 140V for 1.5 hours. Theseparated proteins in the gels are transferred onto PVDF membranes at100V for about 1 hour in Western transfer buffer. The blots areincubated in blocking buffer at room temperature for 1 hour or at 4° C.overnight with gentle rocking. Primary antibodies against various HSP90client proteins of interest are applied at room temperature for 1 hourwith gentle rocking. Excess antibodies are washed off with six 5-minutewashes in TBST. The blots are then incubated in BRP-conjugated secondaryantibodies and streptavidin-HRP conjugate at room temperature for 1 hourwith gentle rocking. Excess secondary antibodies and conjugate arewashed off by six 5 minute washes in TBST. The blots are then developedusing Pierce's SuperSignal West Femto chemilluminescent substrate bymixing freshly prepared equal volumes of luminal enhancer and peroxidebuffer and adding the mixture onto the blots one at a time. The proteinbands can be visualized on Bio-Rad's fluor-S Max2 MultiImager usingBio-Rad's Quantity One software.

The results of the Western Blott analysis are shown in FIG. 5.

The foregoing examples are not limiting and are merely illustrative ofvarious aspects and embodiments of the present invention. All documentscited herein are indicative of the levels of skill in the art to whichthe invention pertains and are incorporated by reference herein in theirentireties. None, however, is admitted to be prior art.

One skilled in the art will readily appreciate that the presentinvention is well adapted to carry out the objects and obtain the endsand advantages mentioned, as well as those inherent therein. The methodsand compositions described illustrate preferred embodiments, areexemplary, and are not intended as limitations on the scope of theinvention. Certain modifications and other uses will occur to thoseskilled in the art, and are encompassed within the spirit of theinvention, as defined by the scope of the claims.

The invention illustratively described herein suitably may be practicedin the absence of any element or elements, limitation or limitationswhich is not specifically disclosed herein. The terms and expressionswhich have been employed are used as terms of description and not oflimitation, and there is no intention in the use of such terms andexpressions of excluding any equivalents of the features shown anddescribed, or portions thereof. It is recognized that variousmodifications are possible within the scope of the invention claimed.Thus, it should be understood that although the present invention hasbeen specifically disclosed by preferred embodiments, optional features,modifications and variations of the concepts herein disclosed may beresorted to by those skilled in the art, and that such modifications andvariations are considered to be within the scope of this invention asdefined by the description and the appended claims.

In addition, where features or aspects of the invention are described interms of Markush groups or other grouping of alternatives, e.g.,genuses, those skilled in the art will recognize that the invention isalso thereby described in terms of any individual member or subgroup ofmembers of the Markush group or subgenus, and exclusions of individualmembers as appropriate, e.g., by proviso.

Other embodiments are within the following claims.

1. A method of treating an individual having a cancer chosen fromgastric cancer, adrenocortical cancer, melanoma, and colon cancercomprising administering to said individual a pharmaceutical compositioncomprising a pharmaceutically effective amount of a compound of FormulaI, or a pharmaceutically acceptable salt thereof,

wherein: R⁰ is hydrogen; R¹ is chloro; R² is —NH₂; R³ is substitutedlower alkyl wherein the substituent on R³ is selected from —OP(O)(OH)₂and —OR⁸ wherein R⁸ is selected from hydrogen, lower alkyl, and —C(O)R⁹wherein R⁹ is selected from H, lower alkyl, —NR¹⁰R¹⁰ and —OR¹¹, whereineach R¹⁰ is independently selected from hydrogen and lower alkyl, or R¹⁰and R¹⁰ taken together with the N atom to which they are attached forman optionally substituted ring comprising 3-7 ring atoms, wherein, inaddition to said N atom, 0-3 of the ring atoms are heteroatoms selectedfrom O, S and N; and R¹¹ is lower alkyl; R⁴ is lower alkylene; and R⁵ is3,5-dimethyl-4-methoxy-pyridin-2-yl.
 2. The method of claim 1, whereinR³ is —(CH₂)_(n)OH, where n is 1, 2, or
 3. 3. The method of claim 1,wherein R³ is —(CH₂)_(m)C(R¹²)₂(CH₂)_(n)OH, wherein m is 0, 1, or 2; nis 1 or 2; and each R¹² is independently hydrogen or lower alkyl.
 4. Themethod of claim 1, wherein R³ is lower alkyl substituted with the group—OP(O)(OH)₂.
 5. The method of claim 1, wherein R⁴ is —CH₂—.
 6. Themethod of claim 1, wherein the compound of Formula I is selected fromcompounds of the formula:


7. The method of claim 1, wherein the compound of Formula I is of theformula:


8. The method of claim 1, wherein the compound of Formula I is of theformula:


9. The method of claim 1, wherein the compound of Formula I is of theformula:


10. The method of claim 1, wherein said individual has gastric cancer.11. The method of claim 1, wherein said individual has adrenocorticalcancer.
 12. The method of claim 1, wherein said individual has melanoma.13. The method of claim 1, wherein said individual has colon cancer. 14.The method of claim 1, further comprising administering at least onetherapeutic agent selected from cytotoxic agents, anti-angiogenesisagents, and anti-neoplastic agents.
 15. The method of claim 14, whereinthe at least one anti-angiogenesis agent is selected from VEGF receptorinhibitors.
 16. The method of claim 14, wherein the at least oneanti-angiogenesis agent is selected from angiostatin and endostatin. 17.The method of claim 14, wherein the at least one anti-neoplastic agentis selected from alkylating agents, anti-metabolites, epidophyllotoxins,anti-neoplastic enzymes, topoisomerase inhibitors, procarbazines,mitoxantrones, platinum coordination complexes, hormonal/anti-hormonaltherapeutic agents, and haematopoietic growth factors.
 18. The method ofclaim 14, wherein the at least one anti-neoplastic agent is selectedfrom anthracyclines, vinca drugs, mitomycins, bleomycins, cytotoxicnucleosides, epothilones, discodermolide, pteridines, diynenes andpodophyllotoxins.
 19. The method of claim 14, wherein the at least oneanti-neoplastic agent is selected from paclitaxel and gemcitabine. 20.The method of claim 14, wherein the at least one anti-neoplastic agentis selected from carminomycin, daunorubicin, aminopterin, methotrexate,dichloromethotrexate, mitomycin C, porfiromycin, 5-fluorouracil,6-mercaptopurine, cytosine arabinoside, podophyllotoxin, etoposide,etoposide phosphate, teniposide, melphalan, vinblastine, vincristine,leuorosidine, vindesine, leurosine, estramustine, carboplatin,cyclophosphamide, bleomycin, ifosamide, melphalan, hexamethyl melamine,thiotepa, cytarabin, edatrexate trimetrexate, dacarbazine,L-asparaginase, camptothecin, topotecan, bicalutamide, flutamide,leuprolide, interferons, and interleukins.